Small molecule NF-κB inhibitors

ABSTRACT

The present invention provides, inter alia, compounds capable of inhibiting NF-κB. Pharmaceutical compositions containing and methods of using the compounds are also provided herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority to U.S. provisional applicationSer. No. 62/272,408, filed Dec. 29, 2015, the entire contents of whichare incorporated by reference herein.

FIELD OF INVENTION

The present invention provides, inter alia, compounds and pharmaceuticalcompositions capable of inhibiting NF-κB, as well methods of using saidcompounds and compositions.

BACKGROUND OF THE INVENTION

NF-κB/Rel (nuclear factor kappa B) is a family of transcription factorsthat includes p50/p105 (NF-κB1), p52/p100 (NF-κB2), p65 (RelA), c-Rel,and RelB. These molecules can homo- or heterodimerize, and are generallysequestered in the cytoplasm by their inhibitors, IκBs. Upon activation,IκBs are degraded by the 26s proteasome and NF-κB dimers migrate intothe nucleus to perform transcriptional activity.

NF-κB (p50/p65) and c-Rel are regulated by the canonical IKKα/β/γ kinasecomplex pathway, whereas RelB and p52 (NF-κB2) are regulated by analternative pathway via the IKKα/NIK complex. Despite this similarity,each NF-κB family member is distinct with regard to tissue expressionpattern, response to receptor signals, and target gene specificity.These differences are evident from the non-redundant phenotypesexhibited by individual NF-κB/Rel knockout mice. Therefore, therapeuticstargeted to different NF-κB/Rel members are likely to have differentbiological effects and toxicity profiles.

Many receptors and stimuli can activate NF-κB/Rel, including TCR/BCR,TNF receptor superfamily (e.g. CD40, TNFR1, TNFR2, BAFF, APRIL, RANK),IL-1/TLR receptors, and Nod-like receptors, as well as activatingoncogenes (e.g. Src, Ras, LMP-1, Tax, v-FLIP), reactive oxygen radicals,radiation, and chemotherapeutic agents. In response to these stimuli,NF-κB/Rel regulates the expression of cytokines, chemokines, andmolecules that play a role in adhesion, the cell cycle, apoptosis, andangiogenesis. As such, NF-κB/Rel transcription factors are importanttherapeutic targets for many human disorders, including inflammation,autoimmune diseases, and cancer, and small molecule inhibitors ofNF-κB/Rel may be useful as therapeutics for these disorders.

SUMMARY OF THE INVENTION

The present invention relates to compounds capable of inhibitingNF-kB/Rel.

One embodiment of the present invention is a compound. The compound hasa structure of formula (I)

wherein:A, B, C, and D are independently selected from the group consisting ofcarbon and nitrogen;X, Y, and Z are independently selected from the group consisting ofoxygen, sulfur, and NR^(a);R₁, R₂, R₃, and R₄ are independently selected from the group consistingof no atom, hydrogen, halogen, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉ alkynyl,aryl, C₁₋₉ heterocyclic, —OH, —OR^(a), —OR^(a)OR^(b),—OR^(a)OR^(b)OR^(c), —OR^(a)(C═O)R^(b), —O(C═O)R^(a), —O(C═O)OR^(a),—O(C═O)NR^(a)R^(b), cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH,—COR^(a), —COOR^(a), —CONR^(a)R^(b), —CONHCONR^(a)R^(b), —NR^(a)R^(b),—NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(a)R^(b),—CSNHCSNR^(a)R^(b), —SH, —SR^(a), —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(a)R^(b);R₅ is selected from the group consisting of hydrogen, C₁₋₉ alkyl, C₁₋₉alkenyl, C₁₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —R^(a)CO, —R^(a)NHCO,and —R^(a)OCO; andR₆, R^(a), R^(b), and R^(c) are independently selected from the groupconsisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉alkynyl, aryl, and C₁₋₉ heterocyclic,or is a crystalline form, hydrate, or pharmaceutically acceptable saltthereof.

Another embodiment of the present invention is a compound. The compoundis selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

A further embodiment of the present invention is a compound. Thecompound is selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

An additional embodiment of the present invention is a compound capableof inhibiting NF-κB. The compound has the structure of formula (I):

wherein:

-   -   A, B, C, and D are independently selected from the group        consisting of carbon and nitrogen;    -   X, Y, and Z are independently selected from the group consisting        of oxygen, sulfur, and NR^(a);    -   R₁, R₂, R₃, and R₄ are independently selected from the group        consisting of no atom, hydrogen, halogen, C₁₋₉ alkyl, C₁₋₉        alkenyl, C₁₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH, —OR^(a),        —OR^(a)OR^(b), —OR^(a)OR^(b)OR^(c), —O(C═O)R^(a), —O(C═O)OR^(a),        —O(C═O)NR^(a)R^(b), cyano, nitro, CF₃, CHF₂, CH₂F, —CHO, —COOH,        —COR^(a), —COOR^(a), —CONR^(a)R^(b), —CONHCONR^(a)R^(b),        —NR^(a)R^(b), —NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a),        —CSOR^(a), —CSNR^(a)R^(b), —CSNHCSNR^(a)R^(b), —SH, —SR^(a),        —S(C═O)R^(a), —S(C═O)OR^(a), —S(C═O)NR^(a)R^(b);    -   R₅ is selected from the group consisting of hydrogen, C₁₋₉        alkyl, C₁₋₉ alkenyl, C₁₋₉ alkynyl, aryl, C₁₋₉ heterocyclic,        —R^(a)CO, —R^(a)NHCO, and —R^(a)OCO; and    -   R₆, R^(a), R^(b), and R^(c) are independently selected from the        group consisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₁₋₉        alkenyl, C₁₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic,        or is a crystalline form, hydrate, or pharmaceutically        acceptable salt thereof.

A further embodiment of the present invention is a compound capable ofinhibiting NF-κB. The compound is selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

An additional embodiment of the present invention is a compound capableof inhibiting NF-κB. The compound is selected from the group consistingof:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

Another embodiment of the present invention is a pharmaceuticalcomposition. The pharmaceutical composition comprises a pharmaceuticallyacceptable carrier and any of the compounds disclosed herein.

A further embodiment of the present invention is a method of inhibitingNF-κB in a cell. The method comprises contacting the cell with any ofthe compounds disclosed herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a line graph of results from 10 nM CD28RE FITCfluorescence polarization assays. FIG. 1B shows a line graph of coldcompetition with specific (CD28RE) and non-specific (Oct1) oligo. FIG.1C is a dot plot showing distribution of fluorescence polarizationsignals in a representative 384-well plate. FIG. 1D shows a line graphof results from 0.33 nM CD28RE FITC fluorescence polarization assays.

FIG. 2 is a line graph showing Rel/NF-κB inhibition by compound 6 byEMSA.

FIG. 3A is a table showing pharmacokinetic data for compound 13. FIG. 3Bis a line graph showing mean plasma concentration of intravenouscompound 13 in mice over time.

FIG. 4A is a table showing pharmacokinetic data for compound 20. FIG. 4Bis a line graph showing mean plasma concentration of intravenouscompound 20 in mice over time.

FIG. 5A is a table showing pharmacokinetic data for compound 26. FIG. 5Bis a line graph showing mean plasma concentration of intravenouscompound 26 in mice over time.

FIG. 6A is a table showing pharmacokinetic data for compound 42. FIG. 6Bis a line graph showing mean plasma concentration of intravenouscompound 42 in mice over time.

FIG. 7A is a table showing pharmacokinetic data for compound 44. FIG. 7Bis a line graph showing mean plasma concentration of intravenouscompound 44 in mice over time.

FIG. 8A is a table showing pharmacokinetic data for compound 46. FIG. 8Bis a line graph showing mean plasma concentration of intravenouscompound 46 in mice over time.

DETAILED DESCRIPTION OF THE INVENTION

The terminology used herein is for the purpose of describing particularembodiments only and is not intended to be limiting. As used in thespecification and the appended claims, the singular forms “a,” “an,” and“the” include plural referents unless the context clearly dictatesotherwise.

For recitation of numeric ranges herein, each intervening number therebetween with the same degree of precision is explicitly contemplated.For example, for the range of 6-9, the numbers 7 and 8 are contemplatedin addition to 6 and 9, and for the range 6.0-7.0, the numbers 6.0, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitlycontemplated.

The term “about” generally indicates within ±0.5%, 1%, 2%, 5%, or up to±10% of the indicated value. For example, an amount of “about 10 wt %”generally indicates, in its broadest sense, 10 wt %±10%, which indicates9.0-11.0 wt %. The term “about” may alternatively indicate a variationor average in a physical characteristic of a group.

One embodiment of the present invention is a compound. The compound hasthe structure of formula (I):

wherein:A, B, C, and D are independently selected from the group consisting ofcarbon and nitrogen;X, Y, and Z are independently selected from the group consisting ofoxygen, sulfur, and NR^(a);R₁, R₂, R₃, and R₄ are independently selected from the group consistingof no atom, hydrogen, halogen, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉ alkynyl,aryl, C₁₋₉ heterocyclic, —OH, —OR^(a), —OR^(a)OR^(b),—OR^(a)OR^(b)OR^(c), —OR^(a)(C═O)R^(b), —O(C═O)R^(a), —O(C═O)OR^(a),—O(C═O)NR^(a)R^(b), cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH,—COR^(a), —COOR^(a), —CONR^(a)R^(b), —CONHCONR^(a)R^(b), —NR^(a)R^(b),—NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(a)R^(b),—CSNHCSNR^(a)R^(b), —SH, —SR^(a), —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(a)R^(b);R₅ is selected from the group consisting of hydrogen, C₁₋₉ alkyl, C₁₋₉alkenyl, C₁₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —R^(a)CO, —R^(a)NHCO,and —R^(a)OCO; andR₆, R^(a), R^(b), and R^(c) are independently selected from the groupconsisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉alkynyl, aryl, and C₁₋₉ heterocyclic,or is a crystalline form, hydrate, or pharmaceutically acceptable saltthereof.

As used herein, the term “compound” refers to two or more atoms that areconnected by one or more chemical bonds. In the present invention,“chemical bonds” and “bonds” are interchangeable and include, but arenot limited to, covalent bonds, ionic bonds, hydrogen bonds, and van derWaals interactions. Covalent bonds of the present invention includesingle, double, and triple bonds. Compounds of the present inventioninclude, but are not limited to, organic molecules. Atoms that comprisethe compounds of the present invention are “linked” if they areconnected by a chemical bond of the present invention.

Organic compounds of the present invention include linear, branched, andcyclic hydrocarbons with or without functional groups. The term“C_(x-y)” when used in conjunction with a chemical moiety, such as,alkyl, alkenyl, alkynyl or alkoxy is meant to include groups thatcontain from x to y carbons in the chain. For example, the term “C_(x-y)alkyl” means substituted or unsubstituted saturated hydrocarbon groups,including straight-chain alkyl and branched-chain alkyl groups thatcontain from x to y carbons in the chain, including haloalkyl groupssuch as trifluoromethyl and 2,2,2-trifluoroethyl, etc. The terms“C_(x-y) alkenyl” and “C_(x-y) alkynyl” refer to substituted orunsubstituted unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but containing atleast one double or triple bond respectively.

The term “independently selected” and grammatical variations thereofmean that, in a chemical structure of the present invention, (e.g.,formula I), if more than one atom in the structure can be selected froma list of elements, those atoms may or may not be of the same element.Similarly, if more than one chemical moiety in the structure can beselected from a list of chemical moieties, those moieties may or may notbe the same.

In one aspect of this embodiment,

X, Y, and Z are independently selected from the group consisting ofoxygen and sulfur;

R₁ is selected from the group consisting of —H, —F, —Cl, —OMe, and —OEt;

R₂ is selected from the group consisting of —H, —CH₃, —OH, —OMe, —OEt,-Me, -Et, -nPr, —O-nPr, —OEtnPr, —OC₄H₉, —OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅,—O-isobutyl, —O-isopentyl, —OC_(n)H_(2n)OMe,—OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

R₃ is selected from the group consisting of —H, —Cl, —Br, —F, and —OMe;R₄ is selected from the group consisting of —H and —OMe;R₅ is selected from the group consisting of —H, -Me, -Et, —Pr, -iPr,-Ph, iBu and -nBu;R₆ is selected from the group consisting of —H and —CH₃;m is 2, 3, 4 or 5; and, n is 2, 3, 4, or 5.

In another aspect of this embodiment,

X, Y, and Z are independently selected from the group consisting ofoxygen and sulfur; and

R₆ is hydrogen.

Preferably,

Z is oxygen;

R₁ and R₃ are selected from the group consisting of hydrogen, halogen,—CN, and —CF₃;

R₂ is selected from the group consisting of C₁₋₉ alkoxy,—OC_(n)H_(2n)OMe, —OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe, and —OH;

R₄ is selected from the group consisting of hydrogen, C₁₋₉ alkoxy and—OH;

m is 2, 3, 4 or 5; and n is 2, 3, 4, or 5.

More preferably,

X and Y are oxygen; and

R₄ is hydrogen.

In an exemplary aspect of this embodiment, the compound is selected fromthe group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

In an additional aspect of this embodiment, R₁ and R₂ are linked by atleast one bond.

In a further aspect of this embodiment, R₂ and R₃ are linked by at leastone bond.

The compound according to claim 1, wherein R₃ and R₄ are linked by atleast one bond.

Another embodiment of the present invention is a compound. The compoundis selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

A further embodiment of the present invention is a compound. Thecompound is selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

An additional embodiment of the present invention is a compound capableof inhibiting NF-κB. The compound has the structure of formula (I):

wherein:A, B, C, and D are independently selected from the group consisting ofcarbon and nitrogen;X, Y, and Z are independently selected from the group consisting ofoxygen, sulfur, and NR^(a);R₁, R₂, R₃, and R₄ are independently selected from the group consistingof no atom, hydrogen, halogen, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉ alkynyl,aryl, C₁₋₉ heterocyclic, —OH, —OR^(a), —OR^(a)OR^(b),—OR^(a)OR^(b)OR^(c), —O(C═O)R^(a), —O(C═O)OR^(a), —O(C═O)NR^(a)R^(b),cyano, nitro, CF₃, CHF₂, CH₂F, —CHO, —COOH, —COR^(a), —COOR^(a),—CONR^(a)R^(b), —CONHCONR^(a)R^(b), —NR^(a)R^(b), —NHCOR^(a),—NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(a)R^(b),—CSNHCSNR^(a)R^(b), —SH, —SR^(a), —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(a)R^(b);R₅ is selected from the group consisting of hydrogen, C₁₋₉ alkyl, C₁₋₉alkenyl, C₁₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —R^(a)CO, —R^(a)NHCO,and —R^(a)OCO; andR₆, R^(a), R^(b), and R^(c) are independently selected from the groupconsisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₁₋₉ alkenyl, C₁₋₉alkynyl, aryl, and C₁₋₉ heterocyclic,or is a crystalline form, hydrate, or pharmaceutically acceptable saltthereof.

In the present invention, the compound capable of inhibiting NF-κB mayfunction as a direct or indirect NF-κB/Rel inhibitor. A direct NF-κB/Relinhibitor is a compound that binds to or interacts with NF-κB/Reldirectly and inhibits its DNA binding and transcriptional function. Anindirect NF-κB/Rel inhibitor is a compound that binds to or interactswith a compound other than NF-κB/Rel, thereby generating a downstreaminhibitory effect on NF-κB/Rel activity.

In one aspect of this embodiment,

X, Y, and Z are independently selected from the group consisting ofoxygen and sulfur;

R₁ is selected from the group consisting of —H, —F, —Cl, —OMe, and —OEt;

R₂ is selected from the group consisting of —H, —CH₃, —OH, —OMe, —OEt,-Et, -nPr, —O-nPr, —OEtnPr, —OC₄H₉, —OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅,—O-isobutyl, —O-isopentyl, —OC_(n)H_(2n)OMe,—OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

R₃ is selected from the group consisting of —H, —Cl, —Br, —F, and —OMe;R₄ is selected from the group consisting of —H and —OMe;R₅ is selected from the group consisting of —H, -Me, and -nBu;R₆ is selected from the group consisting of —H and —CH₃;m is 2, 3, 4 or 5; and n is 2, 3, 4, or 5.

In another aspect of this embodiment,

X, Y, and Z are independently selected from the group consisting ofoxygen and sulfur; and

R₆ is hydrogen.

Preferably,

Z is oxygen;

R₁ and R₃ are selected from the group consisting of hydrogen, halogen,—CN, and —CF₃;

R₂ is selected from the group consisting of C₁₋₉ alkoxy,—OC_(n)H_(2n)OMe, —OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, and —OH;

R₄ is selected from the group consisting of hydrogen, C₁₋₉ alkoxy and—OH;

m is 2, 3, 4 or 5; and n is 2, 3, 4, or 5.

More preferably,

X and Y are oxygen; and

R₄ is hydrogen.

In an exemplary aspect of this embodiment, the compound is selected fromthe group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

In an additional aspect of this embodiment, R₁ and R₂ are linked by atleast one bond.

In a further aspect of this embodiment, R₂ and R₃ are linked by a bond.

In another aspect of this embodiment, R₃ and R₄ are linked by a bond.

A further embodiment of the present invention is a compound capable ofinhibiting NF-κB. The compound is selected from the group consisting of:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

An additional embodiment of the present invention is a compound capableof inhibiting NF-κB. The compound is selected from the group consistingof:

and crystalline forms, hydrates, or pharmaceutically acceptable saltsthereof.

R₁ and R₂, R₂ and R₃, and R₃ and R₄, can be joined to form a ring,aromatic or not. The ring can be a hydrocarbon ring or a heterocyclicring.

Another embodiment of the present invention is a pharmaceuticalcomposition. The pharmaceutical composition comprises a pharmaceuticallyacceptable carrier and any of the compounds disclosed herein. Thepharmaceutical composition may also include any number of otherauxiliary agents used in the art, e.g., buffering agents, stabilizingagents, emulsifying agents, pH adjusting agents, surfactants, andflavorants.

A further embodiment of the present invention is a method of inhibitingNF-κB in a cell. The method comprises contacting the cell with any ofthe compounds disclosed herein.

As used herein, the term “contacting” means bringing a compound of thepresent invention into close proximity to the cells of the presentinvention. This may be accomplished using conventional techniques ofdrug delivery to mammals (e.g., tail vein injection, intravenousinjection, peroral administration) or in the in vitro situation by,e.g., providing a compound of the present invention to a culture mediato which the cells of the present invention are exposed.

Cells of the present invention include any cell type, cancerous ornon-cancerous, in vitro or in vivo, that expresses NF-κB or any NF-κBfamily member. Cells of the present invention include, but are notlimited to, human, monkey, ape, hamster, rat, or mouse cells. In someembodiments, cells of the present invention include, but are not limitedto, CHO (e.g., CHO K1, DXB-11 CHO, Veggie-CHO), COS (e.g., COS-7),retinal cells, Vero, CV1, kidney (e.g., HEK293, 293 EBNA, MSR293, MDCK,HaK, BHK), HeLa, HepG2, WI38, MRC 5, Col0205, HB 8065, HL-60, (e.g.,BHK21), Jurkat, Daudi, A431 (epidermal), CV-1, U937, 3T3, L cells, C127cells, SP2/0, NS-0, MMT 060562, Sertoli cells, BRL 3A cells, HT1080cells, myeloma cells, tumor cells, and any cell line derived from any ofthe aforementioned cells.

DEFINITIONS

The term “aliphatic”, as used herein, means a group composed of carbonand hydrogen atoms that does not contain aromatic rings. Accordingly,aliphatic groups include alkyl, alkenyl, alkynyl, and carbocyclylgroups.

The term “alkyl” means the radical of saturated aliphatic groups thatdoes not have a ring structure, including straight chain alkyl groups,and branched chain alkyl groups. Alkyl groups of the present inventionhave at least one and up to twenty carbon atoms and can be optionallysubstituted with one or more heteroatoms selected from halogen,nitrogen, oxygen, and sulfur.

The term “alkyl” also refers to cyclic hydrocarbon rings having at leastthree, and up to twenty, carbon atoms, optionally substituted with oneor more heteroatoms selected from halogen, nitrogen, oxygen, and sulfur.The cyclic hydrocarbon ring can be monocyclic, bicyclic, polycyclic orbridge cyclic. Examples of alkyl groups include, but are not limited to,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, neopentyl,cyclopropyl, cyclobutyl, 2-chlorobutyl, 3-fluoropentyl, 4-hydroxybutyl,3-methoxypropyl, 2-methoxypropyl, 2-methylbutyl, 3-methylbutyl(isopentyl), 2-chloro-4-hydroxybutyl, 5-aminohexyl,2,2-difluorocyclobutyl, 1,3-difluorocyclohexyl, 3-thiolhexyl, and thelike.

The term “alkenyl” refers to a straight or branched hydrocarbon chainhaving at least three, and up to twenty, carbon atoms with at least onedouble bond (—C═C—), optionally substituted with one or more heteroatomsselected from halogen, nitrogen, oxygen, and sulfur.

The term “alkenyl” also refers to cyclic hydrocarbon rings having atleast three, and up to twenty, carbon atoms with at least one doublebond (—C═C—), optionally substituted with one or more heteroatomsselected from halogen, nitrogen, oxygen, and sulfur. Examples of alkenylgroups include, but are not limited to, ethenyl, chlorovinyl, propenyl,propenylene, allyl, 1,4-butandienyl, 1,2-cyclobutenyl, and the like.

The term “alkynyl” refers to a straight or branched hydrocarbon chainhaving at least three, and up to twenty, carbon atoms with at least onetriple bond, with or without one or more double bond, and optionallysubstituted with one or more heteroatoms selected from halogen,nitrogen, oxygen, and sulfur. The term “alkenyl” also refers to cyclichydrocarbon rings with at least one triple bond with or without one ormore double bond (—C═C—) and optionally substituted with one or moreheteroatoms selected from halogen, nitrogen, oxygen, and sulfur.Examples of alkynyl groups include, but are not limited to, ethynyl,propynyl, butynyl, 3-methylbutynyl, and the like.

The term “aryl” refers to monocyclic, bicyclic or polycyclic aromatichydrocarbon ring structures optionally substituted with one or moreheteroatoms selected from halogen, nitrogen, oxygen, and sulfur, and/oroptionally substituted with one or more alkyl, alkenyl or alkynylgroups. Examples of aryl groups include, but are not limited to, phenyl,naphthyl, hydroxyl phenyl, chlorophenyl, 2-chloro-4-fluorophenyl,methylphenyl, cyanonaphthyl, and the like.

The term “heterocyclic” refers to saturated or unsaturated mono- orpoly-carbocyclic structures in which at least one carbon atom of atleast one of the rings is replaced by nitrogen, sulfur, phosphorus, oroxygen. The term “heterocyclic” is intended to encompass fully saturatedand unsaturated ring systems as well as partially unsaturated ringsystems, including all possible isomeric forms of the heterocycle (forexample, pyrrolyl comprises 1H-pyrrolyl and 2H-pyrrolyl).

Examples of a monocyclic heterocycle (e.g., a 4-, 5-, or 6-memberedring) or a bicyclic (e.g., a 5/6, 5/5, 6/6 system) saturated heterocycleinclude, but are not limited to, tetrahydrofuranyl, pyrrolidinyl,tetrahydrothienyl, dihydrooxazolyl, piperidinyl, hexahydropyrimidinyl,dioxanyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl and thelike.

Examples of a partially saturated monocyclic, bicyclic or tricyclicheterocycle include, but are not limited to, pyrrolinyl, imidazolinyl,pyrazolinyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolanyl,2,3dihydro-1,4-benzodioxinyl, indolinyl and the like.

Examples of an aromatic monocyclic, bicyclic or tricyclic heterocycleinclude, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl,1,2,3-triazolyl, 1,2,5-thiadiazolyl, 1,2,3-thiadiazolyl,1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazinyl, benzofuranyl, isobenzofuranyl, benzothienyl,isobenzothienyl, indolizinyl, indolyl, isoindoly, benzoxazolyl,benzimidazolyl, indazolyl, benzisoxazolyl, benzisothiazolyl,benzopyrazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl,quinolinyl, isoquinolinyl, cinnolinyl, quinolizinyl, phthalazinyl,quinoxalinyl, quinazolinyl, naphthyridinyl, pteridinyl,pyrrolopyridinyl, thienopyridinyl, furanopyridinyl,isothiazolopyridinyl, thiazolopyridinyl, isoxazolopyridinyl,oxazolopyridinyl, pyrazolopyridinyl, imidazopyridinyl, pyrrolopyrazinyl,thienopyrazinyl, furanopyrazinyl, isothiazolopyrazinyl,thiazolopyrazinyl, isoxazolopyrazinyl, oxazolopyrazinyl,pyrazolopyrazinyl, imidazopyrazinyl, pyrrolopyrimidinyl,thienopyrimidinyl, furanopyrimidinyl, isothiazolopyrimidinyl,thiazolopyrimidinyl, isoxazolopyrimidinyl, oxazolopyrimidinyl,pyrazolopyrimidinyl, imidazopyrimidinyl, pyrrolopyridazinyl,thienopyridazinyl, furanopyridazinyl, isothiazolopyridazinyl,thiazolopyridazinyl, isoxazolopyridazinyl, oxazolopyridazinyl,pyrazolopyridazinyl, imidazopyridazinyl, oxadiazolopyridinyl,thiadiazolopyridinyl, triazolopyridinyl, oxadiazolopyrazinyl,thiadiazolopyrazinyl, triazolopyrazinyl, oxadiazolopyrimidinyl,thiadiazolopyrim idinyl, triazolopyrimidinyl, oxadiazolopyridazinyl,thiadiazolopyridazinyl, triazolopyridazinyl, isoxazolotriazinyl,isothiazolotriazinyl, pyrazolotriazinyl, oxazolotriazinyl,thiazolotriazinyl, imidazotriazinyl, oxadiazolotriazinyl,thiadiazolotriazinyl, triazolotriazinyl, carbazolyl and the like.

The term “carbonyl” includes, but is not limited to, CHO (aldehydegroup), COOH (carboxylic acid), COR^(a) (ketone), COOR^(a) (carboxylicester), CONR^(a)R^(b) (amide), CONHCONR^(a)R^(b) (imide), R^(a)COX (acylhalide), and R^(a)COOCOR^(b) (acid anhydride).

The term “halogen” includes, but is not limited to, fluorine, chlorine,bromine, iodine, and astatine.

In the present invention, the term “crystalline form” means the crystalstructure of a compound. A compound may exist in one or more crystallineforms, which may have different structural, physical, pharmacological,or chemical characteristics. Different crystalline forms may be obtainedusing variations in nucleation, growth kinetics, agglomeration, andbreakage.

Nucleation results when the phase-transition energy barrier is overcome,thereby allowing a particle to form from a supersaturated solution.Crystal growth is the enlargement of crystal particles caused bydeposition of the chemical compound on an existing surface of thecrystal. The relative rate of nucleation and growth determine the sizedistribution of the crystals that are formed. The thermodynamic drivingforce for both nucleation and growth is supersaturation, which isdefined as the deviation from thermodynamic equilibrium. Agglomerationis the formation of larger particles through two or more particles(e.g., crystals) sticking together and forming a larger crystallinestructure.

The term “hydrate”, as used herein, means a solid or a semi-solid formof a chemical compound containing water in a molecular complex. Thewater is generally in a stoichiometric amount with respect to thechemical compound.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the compounds disclosed herein wherein the compounds are modified bymaking acid or base salts thereof. Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like. For example,such salts include salts from ammonia, L-arginine, betaine, benethamine,benzathine, calcium hydroxide, choline, deanol, diethanolamine(2,2′-iminobis(ethanol)), diethylamine, 2-(diethylamino)-ethanol,2-aminoethanol, ethylenediamine, N-ethyl-glucamine, hydrabamine,1H-imidazole, lysine, magnesium hydroxide,4-(2-hydroxyethyl)-morpholine, piperazine, potassium hydroxide,1-(2-hydroxy-ethyl)-pyrrolidine, sodium hydroxide, triethanolamine(2,2′,2″-nitrilotris(ethanol)), trometh-amine, zinc hydroxide, aceticacid, 2.2-dichloro-acetic acid, adipic acid, alginic acid, ascorbicacid, L-aspartic acid, benzenesulfonic acid, benzoic acid,2,5-dihydroxybenzoic acid, 4-acetamido-benzoic acid, (+)-camphoric acid,(+)-camphor-10-sulfonic acid, carbonic acid, cinnamic acid, citric acid,cyclamic acid, decanoic acid, dodecylsulfuric acid,ethane-1,2-disulfonic acid, ethanesulfonic acid,2-hydroxy-ethanesulfonic acid, ethylenediamonotetraacetic acid, formicacid, fumaric acid, galacaric acid, gentisic acid, D-glucoheptonic acid,D-gluconic acid, D-glucuronic acid, glutamic acid, glutantic acid,glutaric acid, 2-oxo-glutaric acid, glycero-phosphoric acid, glycine,glycolic acid, hexanoic acid, hippuric acid, hydrobromic acid,hydrochloric acid isobutyric acid, DL-lactic acid, lactobionic acid,lauric acid, lysine, maleic acid, (−)-L-malic acid, malonic acid,DL-mandelic acid, methanesulfonic acid, galactaric acid,naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid,1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, octanoic acid,oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid(embonic acid), phosphoric acid, propionic acid, (−)-L-pyroglutamicacid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearicacid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid,thiocyanic acid, p-toluenesulfonic acid and undecylenic acid. Furtherpharmaceutically acceptable salts can be formed with cations from metalslike aluminum, calcium, lithium, magnesium, potassium, sodium, zinc andthe like. (Pharmaceutical salts, Berge, S. M. et al., J. Pharm. Sci.,(1977), 66, 1-19).

The pharmaceutically acceptable salts of the present invention can besynthesized from a compound disclosed herein which contains a basic oracidic moiety by conventional chemical methods. Generally, such saltscan be prepared by reacting the free acid or base forms of thesecompounds with a sufficient amount of the appropriate base or acid inwater or in an organic diluent like ether, ethyl acetate, ethanol,isopropanol, or acetonitrile, or a mixture thereof.

The compositions of the invention comprise one or more activeingredients in admixture with one or more pharmaceutically acceptablediluents or carriers and, optionally, one or more other compounds,drugs, ingredients and/or materials. Regardless of the route ofadministration selected, the agents/compounds of the present inventionare formulated into pharmaceutically-acceptable dosage forms byconventional methods known to those of skill in the art. See, e.g.,Remington, The Science and Practice of Pharmacy (21 st Edition,Lippincott Williams and Wilkins, Philadelphia, Pa.).

Pharmaceutically acceptable diluents or carriers are well known in theart (see, e.g., Remington, The Science and Practice of Pharmacy (21stEdition, Lippincott Williams and Wilkins, Philadelphia, Pa.) and TheNational Formulary (American Pharmaceutical Association, Washington,D.C.)) and include sugars (e.g., lactose, sucrose, mannitol, andsorbitol), starches, cellulose preparations, calcium phosphates (e.g.,dicalcium phosphate, tricalcium phosphate and calcium hydrogenphosphate), sodium citrate, water, aqueous solutions (e.g., saline,sodium chloride injection, Ringer's injection, dextrose injection,dextrose and sodium chloride injection, lactated Ringer's injection),alcohols (e.g., ethyl alcohol, propyl alcohol, and benzyl alcohol),polyols (e.g., glycerol, propylene glycol, and polyethylene glycol),organic esters (e.g., ethyl oleate and tryglycerides), biodegradablepolymers (e.g., polylactide-polyglycolide, poly(orthoesters), andpoly(anhydrides)), elastomeric matrices, liposomes, microspheres, oils(e.g., corn, germ, olive, castor, sesame, cottonseed, and groundnut),cocoa butter, waxes (e.g., suppository waxes), paraffins, silicones,talc, silicylate, etc. Each pharmaceutically acceptable diluent orcarrier used in a pharmaceutical composition of the invention must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not injurious to the subject. Diluents orcarriers suitable for a selected dosage form and intended route ofadministration are well known in the art, and acceptable diluents orcarriers for a chosen dosage form and method of administration can bedetermined using ordinary skill in the art.

The compositions of the invention may, optionally, contain additionalingredients and/or materials commonly used in pharmaceuticalcompositions. These ingredients and materials are well known in the artand include (1) fillers or extenders, such as starches, lactose,sucrose, glucose, mannitol, and silicic acid; (2) binders, such ascarboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,hydroxypropylmethyl cellulose, sucrose and acacia; (3) humectants, suchas glycerol; (4) disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,sodium starch glycolate, cross-linked sodium carboxymethyl cellulose andsodium carbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as cetyl alcohol and glycerol monostearate; (8)absorbents, such as kaolin and bentonite clay; (9) lubricants, such astalc, calcium stearate, magnesium stearate, solid polyethylene glycols,and sodium lauryl sulfate; (10) suspending agents, such as ethoxylatedisostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters,microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agarand tragacanth; (11) buffering agents; (12) excipients, such as lactose,milk sugars, polyethylene glycols, animal and vegetable fats, oils,waxes, paraffins, cocoa butter, starches, tragacanth, cellulosederivatives, polyethylene glycol, silicones, bentonites, silicic acid,talc, salicylate, zinc oxide, aluminum hydroxide, calcium silicates, andpolyamide powder; (13) inert diluents, such as water or other solvents;(14) preservatives; (15) surface-active agents; (16) dispersing agents;(17) control-release or absorption-delaying agents, such ashydroxypropylmethyl cellulose, other polymer matrices, biodegradablepolymers, liposomes, microspheres, aluminum monostearate, gelatin, andwaxes; (18) opacifying agents; (19) adjuvants; (20) wetting agents; (21)emulsifying and suspending agents; (22), solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan; (23)propellants, such as chlorofluorohydrocarbons and volatile unsubstitutedhydrocarbons, such as butane and propane; (24) antioxidants; (25) agentswhich render the formulation isotonic with the blood of the intendedrecipient, such as sugars and sodium chloride; (26) thickening agents;(27) coating materials, such as lecithin; and (28) sweetening,flavoring, coloring, perfuming and preservative agents. Each suchingredient or material must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the subject. Ingredients and materials suitable for aselected dosage form and intended route of administration are well knownin the art, and acceptable ingredients and materials for a chosen dosageform and method of administration may be determined using ordinary skillin the art.

The compositions of the present invention suitable for oraladministration may be in the form of capsules, cachets, pills, tablets,powders, granules, a solution or a suspension in an aqueous ornon-aqueous liquid, an oil-in-water or water-in-oil liquid emulsion, anelixir or syrup, a pastille, a bolus, an electuary or a paste. Theseformulations may be prepared by methods known in the art, e.g., by meansof conventional pan-coating, mixing, granulation or lyophilizationprocesses.

Solid dosage forms for oral administration (capsules, tablets, pills,dragees, powders, granules and the like) may be prepared, e.g., bymixing the active ingredient(s) with one or morepharmaceutically-acceptable diluents or carriers and, optionally, one ormore fillers, extenders, binders, humectants, disintegrating agents,solution retarding agents, absorption accelerators, wetting agents,absorbents, lubricants, and/or coloring agents. Solid compositions of asimilar type may be employed as fillers in soft and hard-filled gelatincapsules using a suitable excipient. A tablet may be made by compressionor molding, optionally with one or more accessory ingredients.Compressed tablets may be prepared using a suitable binder, lubricant,inert diluent, preservative, disintegrant, surface-active or dispersingagent. Molded tablets may be made by molding in a suitable machine. Thetablets, and other solid dosage forms, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient therein.They may be sterilized by, for example, filtration through abacteria-retaining filter. These compositions may also optionallycontain opacifying agents and may be of a composition such that theyrelease the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.The active ingredient can also be in microencapsulated form.

Liquid dosage forms for oral administration includepharmaceutically-acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. The liquid dosage forms may containsuitable inert diluents commonly used in the art. Besides inertdiluents, the oral compositions may also include adjuvants, such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents. Suspensions maycontain suspending agents.

The compositions of the present invention for rectal or vaginaladministration may be presented as a suppository, which may be preparedby mixing one or more active ingredient(s) with one or more suitablenonirritating diluents or carriers which are solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound. The pharmaceuticalcompositions of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such pharmaceutically-acceptablediluents or carriers as are known in the art to be appropriate.

Dosage forms for the topical or transdermal administration includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches, drops and inhalants. The active agent(s)/compound(s) may bemixed under sterile conditions with a suitablepharmaceutically-acceptable diluent or carrier. The ointments, pastes,creams and gels may contain excipients. Powders and sprays may containexcipients and propellants.

The compositions of the present invention suitable for parenteraladministrations may comprise one or more agent(s)/compound(s) incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or non-aqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsuitable antioxidants, buffers, solutes which render the formulationisotonic with the blood of the intended recipient, or suspending orthickening agents. Proper fluidity can be maintained, for example, bythe use of coating materials, by the maintenance of the requiredparticle size in the case of dispersions, and by the use of surfactants.These pharmaceutical compositions may also contain suitable adjuvants,such as wetting agents, emulsifying agents and dispersing agents. It mayalso be desirable to include isotonic agents. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption.

In some cases, in order to prolong the effect of a drug (e.g.,pharmaceutical formulation), it is desirable to slow its absorption fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material havingpoor water solubility.

The rate of absorption of the active agent/drug then depends upon itsrate of dissolution which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of aparenterally-administered agent/drug may be accomplished by dissolvingor suspending the active agent/drug in an oil vehicle. Injectable depotforms may be made by forming microencapsule matrices of the activeingredient in biodegradable polymers. Depending on the ratio of theactive ingredient to polymer, and the nature of the particular polymeremployed, the rate of active ingredient release can be controlled. Depotinjectable formulations are also prepared by entrapping the drug inliposomes or microemulsions which are compatible with body tissue. Theinjectable materials can be sterilized for example, by filtrationthrough a bacterial-retaining filter.

Any formulation of the invention may be presented in unit-dose ormulti-dose sealed containers, for example, ampules and vials, and may bestored in a lyophilized condition requiring only the addition of thesterile liquid diluent or carrier, for example water for injection,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the type described above.

The following examples are provided to further illustrate the methods ofthe present invention. These examples are illustrative only and are notintended to limit the scope of the invention in any way.

EXAMPLES Example 1 General Synthesis Scheme

The NF-κB inhibitors of the present invention can be synthesized by anyof the suitable methods known in the art, or as further described below.

Example 2 Compound-Specific Synthesis Protocols Preparation of9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1: Preparationof compound 1a,5-(2-hydroxy-3-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 1a

In a round bottom flask, a mixture of O-vanillin (1.52 g, 10.0 mmol) andbarbituric acid (1.28 g, 10.0 mmol) in ethanol (20 ml) was stirred andheated to 30° C. overnight. The reaction mixture was cooled to roomtemperature. Subsequently, the reaction mixture was filtered. Solidyellow cake was collected and vacuum dried at 40° C. to 50° C. to yielddesired product 1a, (2.20 g, 8.4 mmol, 84%).

Step 2: Preparation of9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 1a, (500 mg, 1.9 mmol)in a mixture of acetic acid (15 ml) and acetic anhydride (2 ml) wasstirred and heated to 80° C. After 10 minutes at 80° C., the mixtureturned homogeneous. The mixture was stirred at 80° C. for 3 hours. Themixture was cooled to room temperature and stirred overnight.Precipitation occurred during overnight stirring. Yellow solid wasfiltered, washed with water and vacuum dried at 40° C. to 50° C. toyield compound 1 (355 mg, 1.45 mmol, 77%).

Preparation of 9-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of compound 2a,5-(3-ethoxy-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 2a

In a scintillation vial, a mixture of 3-ethoxysalicyladehide (0.332 g,2.0 mmol) and barbituric acid (0.256 g, 2.0 mmol) in ethanol (10 ml) wasstirred and heated to 40° C. overnight. The reaction mixture was cooledto room temperature. Subsequently, the reaction mixture was filtered.Solid yellow cake was collected and vacuum dried at 40° C. to 50° C. toyield desired product 2a, (0.37 g, 1.3 mmol, 67%).

Step 2: Preparation of9-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 2a, (138 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. After 10 minutes at 80° C., the mixtureturned homogeneous and remained homogeneous during the heating. Themixture was stirred at 80° C. for 1 hour. The mixture was cooled to roomtemperature. Acetic acid and acetic anhydride were removed from thereaction mixture by N₂ stream blowing to cause precipitation. Yellowsolid was filtered, washed with water and vacuum dried at 40° C. to 50°C. to yield compound 2 (62 mg, 0.24 mmol, 48%).

Preparation of7-chloro-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of5-(5-chloro-2-hydroxy-3-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,3a

In a scintillation vial, a mixture of5-chloro-2-hydroxy-3-methoxybenzaldehyde (0.374 g, 2.0 mmol) andbarbituric acid (0.256 g, 2.0 mmol) in a mixture of ethanol (10 ml) andwater (5 ml) was stirred and heated to 25° C. for 2 hours. The reactionmixture was cooled to room temperature. Subsequently, the reactionmixture was filtered. Solid yellow cake was collected and vacuum driedat 40° C. to 50° C. to yield desired product 3a, (0.51 g, 1.7 mmol,86%).

Step 2: Preparation of7-chloro-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 3a, (148 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 1.5 hour. The mixture was cooled toroom temperature. Acetic acid and acetic anhydride were removed from thereaction mixture by N₂ stream blowing to cause precipitation. Yellowsolid was filtered, washed with water and vacuum dried at 40° C. to 50°C. to yield compound 3 (98 mg, 0.35 mmol, 70%).

Preparation of7-bromo-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of5-(5-bromo-2-hydroxy-3-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,4a

A solution of 5-bromo-2-hydroxy-3-methoxybenzaldehyde (0.462 g, 2.0mmol) in ethanol (40 ml) was added to a solution of barbituric acid(0.256 g, 2.0 mmol) in water (40 ml). The mixture was stirred at roomtemperature for 2 days. Subsequently, the reaction mixture was filtered.Orange solid cake was collected and vacuum dried at 40° C. to 50° C. toyield desired product 4a, (0.46 g, 1.34 mmol, 67%).

Step 2: Preparation of7-bromo-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 4a, (170 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 30 hours. The mixture was cooled toroom temperature. Acetic acid and acetic anhydride were removed from thereaction mixture by N₂ stream blowing to cause precipitation. Yellowsolid was filtered, washed with water and vacuum dried at 40° C. to 50°C. to yield compound 4 (145 mg, 0.45 mmol, 90%).

Preparation of7-chloro-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 5-chloro-3-ethoxy-2-hydroxybenzaldehyde, 5a

In a round bottom flask, N-chlorosuccinimide (8.68 g, 65 mmol) was addedto a solution of 3-ethoxysalicylaldehyde (8.30 g, 50 mmol) in THF (100ml). The mixture was stirred at room temperature for 5 hours. Thereaction was quenched with water (100 ml) and extracted with ethylacetate (150 ml). The organic layer was washed successively withsaturated NaHCO₃ aq, HCl aq. (0.5 M, 150 ml) and then with brine (150ml). Organic layer was concentrated to dryness to yield compound 5a(9.60 g, 48 mmol, 96%) that can be used in the next step without furtherpurification.

Step 2: Preparation of5-(5-bromo-2-hydroxy-3-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,5b

In a scintillation vial, a mixture of 5a (0.40 g, 2.0 mmol), barbituricacid (0.256 g, 2.0 mmol), ethanol (7 ml) and water (10 ml) was stirredat room temperature for 2 hours. Subsequently, the reaction mixture wasfiltered. Solid yellow cake was collected and vacuum dried at 40° C. to50° C. to yield desired product 5b, (0.558 g, 1.88 mmol, 94%).

Step 3: Preparation of7-chloro-9-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 5b, (100 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 0.5 hour. The mixture turnedhomogeneous after 10 minutes at 80° C. Precipitation reoccurred after 20minutes at 80° C. The reaction was stopped after 30 minutes. The mixturewas cooled to room temperature. Yellow solid was filtered, washed withwater and vacuum dried at 40° C. to 50° C. to yield compound 5 (58 mg,0.20 mmol, 40%).

Preparation of 8-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of5-(2-hydroxy-4-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 6a

In a scintillation vial, a mixture of 2-hydroxy-4-methoxybenzaldehyde(0.456 g, 3.0 mmol), barbituric acid (0.384 g, 3.0 mmol), ethanol (10ml) and water (5 ml) was stirred at room temperature for 15 hours.Subsequently, the reaction mixture was filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield desired product6a, (0.695 g, 2.65 mmol, 88%).

Step 2: Preparation of8-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 6a, (278 mg, 1.0 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 2 hours. The mixture turned homogeneousafter 10 minutes at 80° C. The mixture was cooled to room temperature.Yellow solid was filtered, washed with ethanol and then with water andvacuum dried at 40° C. to 50° C. to yield compound 6 (148 mg, 0.60 mmol,60%).

Preparation of7-chloro-9-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one Step1: Preparation of5-(5-chloro-2-hydroxy-3-methoxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,7a

In a scintillation vial, a mixture of5-chloro-2-hydroxy-3-methoxybenzaldehyde (0.374 g, 2.0 mmol),2-thiobarbituric acid (0.288 g, 3.0 mmol), ethanol (15 ml) and water (5ml) was stirred at room temperature for 15 hours. Subsequently, thereaction mixture was filtered. Solid orange cake was collected andvacuum dried at 40° C. to 50° C. to yield desired product 7a, (0.47 g,1.5 mmol, 75%).

Step 2: Preparation of7-chloro-9-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 7a, (158 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 2 hours. The mixture was cooled to roomtemperature. Red solid was filtered, washed with ethanol and then withwater and vacuum dried at 40° C. to 50° C. to yield compound 7 (62 mg,0.21 mmol, 42%).

Preparation of 8-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-oneStep 1: Preparation of5-(2-hydroxy-4-methoxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,8a

In a scintillation vial, a mixture of 2-hydroxy-4-methoxybenzaldehyde(0.456 g, 3.0 mmol), 2-thiobarbituric acid (0.288 g, 3.0 mmol), ethanol(10 ml) and water (5 ml) was stirred at room temperature for 2 hours.Subsequently, the reaction mixture was filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield desired product8a, (0.761 g, 2.73 mmol, 91%).

Step 2: Preparation of8-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 8a, (150 mg, 0.54mmol) in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml)was stirred and heated to 80° C. for 2 hours. The mixture was cooled toroom temperature. Red solid was filtered, and re-suspended in saturatedaqueous solution of NaHCO3 for 1 hour. Red solid was filtered again,washed with water and vacuum dried at 40° C. to 50° C. to yield compound8 (84 mg, 0.32 mmol, 60%).

Preparation of 2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of5-(2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 9a

In a scintillation vial, a mixture of salicylaldehyde (0.122 g, 1.0mmol), barbituric acid (0.128 g, 1.0 mmol) and water (10 ml) was stirredat room temperature for 1 hour. Subsequently, the reaction mixture wasfiltered. Solid yellow cake was collected and vacuum dried at 40° C. to50° C. to yield desired product 9a, (0.761 g, 2.73 mmol, 91%).

Step 2: Preparation of 2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 9a, (116 mg, 0.5 mmol)in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml) wasstirred and heated to 80° C. for 2 hours. The mixture was cooled to roomtemperature. Yellow solid was filtered, washed with water and vacuumdried at 40° C. to 50° C. to yield compound 9 (43 mg, 0.20 mmol, 40%).

Preparation of 7-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of5-(2-hydroxy-5-methoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 10a

In a scintillation vial, a mixture of 2-hydroxy-5-methoxybenzaldehyde(0.304 g, 2.0 mmol), barbituric acid (0.256 g, 2.0 mmol) and water (10ml) was stirred at room temperature for 3 hours. Subsequently, thereaction mixture was filtered. Solid red cake was collected and vacuumdried at 40° C. to 50° C. to yield desired product 10a, (0.42 g, 1.60mmol, 80%).

Step 2: Preparation of7-methoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 10a, (131 mg, 0.5mmol) in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml)was stirred and heated to 80° C. for 3 hours. The mixture was cooled toroom temperature. Yellow solid was filtered, washed with water andvacuum dried at 40° C. to 50° C. to yield compound 10 (110 mg, 0.20mmol, 90%).

Preparation of 8-hydroxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of5-(2,4-dihydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 11a

In a scintillation vial, a mixture of 2,4-dihydroxybenzaldehyde (0.276g, 2.0 mmol), barbituric acid (0.256 g, 2.0 mmol) and ethanol (8 ml) wasstirred at room temperature for 15 hours. Subsequently, the reactionmixture was filtered. Solid orange cake was collected and vacuum driedat 40° C. to 50° C. to yield desired product 11a, (0.447 g, 1.8 mmol,90%).

Step 2: Preparation of8-hydroxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 11a, (124 mg, 0.5mmol) in a mixture of acetic acid (3 ml) and acetic anhydride (1 ml) wasstirred and heated to 80° C. for 18 hours. The mixture was cooled toroom temperature and stirred overnight. Yellow solid was filtered,washed with water and vacuum dried at 40° C. to 50° C. to yield compound11 (85 mg, 0.37 mmol, 74%).

Preparation of 8-methyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of5-(2-hydroxy-4-methylbenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 12a

In a scintillation vial, a mixture of 2-hydroxy-4-methylbenzaldehyde(0.272 g, 2.0 mmol), barbituric acid (0.256 g, 2.0 mmol) and water (10ml) was stirred at room temperature for 3 hours. Subsequently, ethylacetate (5 ml) was added to the reaction mixture to wash unreacted2-hydroxy-4-methylbenzaldehyde. The reaction mixture was filtered. Solidyellow cake was collected and vacuum dried at 40° C. to 50° C. to yielddesired product 12a, (0.461 g, 1.87 mmol, 94%).

Step 2: Preparation of8-methyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 12a, (123 mg, 0.5mmol) in a mixture of acetic acid (2.5 ml) and acetic anhydride (0.25ml) was stirred and heated to 80° C. for 5 hours. The mixture was cooledto room temperature and stirred overnight. Yellow solid was filtered,washed with water and vacuum dried at 40° C. to 50° C. to yield compound12 (100 mg, 0.44 mmol, 88%).

Preparation of 8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of 4-ethoxy-2-hydroxybenzaldehyde, 13a

A suspension of 2,4-dihydroxybenzaldehyde (3.04 g, 20 mmol), ethylbromide (2.29 g, 21 mmol), potassium carbonate (2.90 g, 21 mmol), NaI(2.98 g, 20 mmol) and 18-crown-6 (0.528 g, 2 mmol) in acetone (15 ml)was stirred and heated to 70° C. for 15 hours. The mixture was cooled toroom temperature and quenched with HCl, aq. 1M. The mixture wasextracted with ethyl acetate (50 ml). Organic layer was washed withwater (50 ml) twice and then with brine (50 ml). Organic layer was driedover MgSO4, concentrated and chromatographed to yield4-ethoxy-2-hydroxybenzaldehyde (3.01 g, 18.1 mmol, 90%) and2,4-diethoxybenzaldehyde (0.134 g, 0.7 mmol, 3.4%).

Step 2: Preparation of5-(4-ethoxy-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 13b

In a scintillation vial, a mixture of 2-hydroxy-4-ethoxybenzaldehyde(0.498 g, 3.0 mmol), barbituric acid (0.384 g, 3.0 mmol) and water (10ml) was stirred at room temperature for 2 hours. The reaction mixturewas filtered. Solid yellow cake was collected and vacuum dried at 40° C.to 50° C. to yield desired product 13b, (0.68 g, 2.46 mmol, 82%).

Step 3: Preparation of8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 13b, (276 mg, 1.0mmol) in a mixture of acetic acid (5.0 g) and acetic anhydride (0.612 g,6 mmol) was stirred and heated to 80° C. for 3 hours. Yellow solid wasfiltered, washed with water and vacuum dried at 40° C. to 50° C. toyield compound 13 (177 mg, 0.68 mmol, 68%).

Preparation of 6,8-dimethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1: Preparation of 2,4,6-trimethoxybenzaldehyde, 14a

In a round bottom flask, POCl3 (7.3 ml, 80 mmol) was added to a mixtureof 1,3,5-trimethoxybenzene (8.41 g, 50 mmol) in DMF (15 ml). Thetemperature of reaction mixture was kept below 30° C. by slowly addingPOCl3. After addition of POCl3, the mixture was still stirred at roomtemperature for an addition 1 hour. The reaction mixture was added intoa cold saturated NaHCO3 solution. The pH of the solution was adjusted toremain above 7. Precipitation of desired product occurred. The off-whiteproduct was filtered and rinsed with NaHCO3 aq., then with HCl aq. 0.5Mand then with water. Off-white solid was collected and vacuum dried atroom temperature to yield 14a (9.60 g, 48 mmol, 98%).

Step 2: 2-hydroxy-4,6-dimethoxybenzaldehyde, 14b

A solution of 14a (5.88 g, 30 mmol) in dichloromethane (40 ml) in around bottom flask was cooled to 0° C. (ice+brine bath). To the solutionwas added BCl3 (45 ml of 1M solution in hexane, 45 mmol). The reactionmixture was allowed to slowly warm to room temperature over 1 hour.After 1.5 hour of reaction, TLC analysis indicated that trace amount ofstarting material remained. An additional BCl₃ (10 ml of 1M solution inhexane, 10 mmol) was added into the mixture. The mixture was stirred atroom temperature for an additional 1 hour. Subsequently, the reactionmixture was quenched with HCl aq. 1M (100 ml). The mixture was extractedwith ethyl acetate. The organic layer was successively washed with HClaq. 1M and brine, and then dried over Mg₂SO₄. The organic layer wasconcentrated and chromatographed to yield 14b (5.44 g, 29.8 mmol, 99%).

Step 3: Preparation of5-(2-hydroxy-4,6-dimethoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,14c

In a scintillation vial, a mixture of 4,6-dimethoxysalicylaldehyde(0.364 g, 2.0 mmol), barbituric acid (0.256 g, 2.0 mmol) and water (10ml) was stirred at room temperature for 4.5 hours. The reaction mixturewas filtered. Solid orange cake was collected and vacuum dried at 40° C.to 50° C. to yield a mixture containing 4,6-dimethoxysalicylaldehyde,5-(2-hydroxy-4,6-dimethoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trioneand5-(6,8-dimethoxy-2,4-dioxo-2,3,4,5-tetrahydro-1H-chromeno[2,3-d]pyrimidin-5-yl)pyrimidine-2,4,6(1H,3H,5H)-trione.The mixture was purified by (1) washed with hot ethyl acetate to remove4,6-dimethoxysalicylaldehyde; (2) re suspended solid obtained afterethyl acetate wash in NaHCO₃ aq. The suspension was filtered to obtain ayellow cake. The yellow cake was vacuum dried at 40° C. to 50° C. toyield desired product 14c, (56 mg, 0.19 mmol, 9.6%).

Step 4: Preparation of6,8-dimethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 14c, (56 mg, 0.19mmol) in a mixture of acetic acid (2 ml) and acetic anhydride (0.2 ml)was stirred and heated to 80° C. for 5 hours. Yellow solid was filtered,washed with water and vacuum dried at 40° C. to 50° C. to yield compound14 (35 mg, 0.13 mmol, 67%).

Preparation of 7-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-oneStep 1: Preparation of5-(2-hydroxy-5-methoxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,15a

In a scintillation vial, a mixture of 2-hydroxy-5-methoxybenzaldehyde(0.456 g, 3.0 mmol), 2-thiobarbituric acid (0.432 g, 2.0 mmol), ethanol(10 ml) and water (5 ml) was stirred at room temperature for 15 hours.Subsequently, the reaction mixture was filtered. Solid red cake wascollected and vacuum dried at 40° C. to 50° C. to yield desired product15a, (0.716 g, 2.57 mmol, 86%).

Step 2: Preparation of7-methoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 15a, (139 mg, 0.5mmol) in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml)was stirred and heated to 80° C. for 4 hours. The mixture was cooled toroom temperature. Bordeaux red solid was filtered, washed with water andvacuum dried at 40° C. to 50° C. to yield compound 15 (85 mg, 0.33 mmol,65%).

Preparation of 8-hydroxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-oneStep 1: Preparation of5-(2,4-dihydroxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,16a

In a scintillation vial, a mixture of 2,4-dihydroxybenzaldehyde (0.414g, 3.0 mmol), 2-thiobarbituric acid (0.432 g, 2.0 mmol), ethanol (10 ml)and water (5 ml) was stirred at room temperature for 5 hours.Subsequently, the reaction mixture was filtered. Solid red cake wascollected and vacuum dried at 40° C. to 50° C. to yield desired product16a, (0.697 g, 2.60 mmol, 88%).

Step 2: Preparation of8-hydroxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 16a, (132 mg, 0.5mmol) in a mixture of acetic acid (4.5 ml) and acetic anhydride (0.5 ml)was stirred and heated to 80° C. for 4 hours. The mixture was cooled toroom temperature. Orange solid was filtered, washed with water andvacuum dried at 40° C. to 50° C. to yield compound 16 (115 mg, 0.47mmol, 93%).

Preparation of 8-methyl-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-oneStep 1: Preparation of5-(2-hydroxy-4-methylbenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,17a

In a scintillation vial, a mixture of 2-hydroxy-4-methoxybenzaldehyde(0.272 g, 3.0 mmol), 2-thiobarbituric acid (0.288 g, 2.0 mmol), andwater (10 ml) was stirred at room temperature for 13 hours. Ethylacetate (5 ml) was added to the reaction mixture to wash unreacted2-hydroxy-4-methoxybenzaldehyde. Subsequently, the reaction mixture wasfiltered. Solid yellow cake was collected and vacuum dried at 40° C. to50° C. to yield desired product 17a, (0.437 g, 1.67 mmol, 83%).

Step 2: Preparation of8-methyl-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 17a, (131 mg, 0.5mmol) in a mixture of acetic acid (2.5 ml) and acetic anhydride (0.25ml) was stirred and heated to 80° C. for 7 hours. The mixture was cooledto room temperature. Yellow solid was filtered, washed with NaHCO3 aq.,then with water and vacuum dried at 40° C. to 50° C. to yield compound17 (33 mg, 0.13 mmol, 27%).

Preparation of6,8-dimethoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one Step 1,Preparation of5-(2-hydroxy-4,6-dimethoxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione,18a

In a scintillation vial, a mixture of 4,6-dimethoxysalicylaldehyde(0.364 g, 2.0 mmol), 2-thiobarbituric acid (0.288 g, 2.0 mmol) and water(15 ml) was stirred and warmed to 50° C. for 30 minutes. The mixture wascooled to room temperature for 15 hours. The reaction mixture wasfiltered. Solid orange cake was collected and re-suspended in ethylacetate (20 ml). The suspension was filtered to remove ethyl acetate andunreacted 4,6-dimethoxysalicylaldehyde. The solid orange from ethylacetate was again re-suspended in saturated NaHCO3 aq. The suspensionwas filtered and washed with water. The solid orange from saturatedNaHCO3 aq. suspension was vacuum dried at 40° C. to 50° C. to yieldcompound 18a, (0.202 g, 0.65 mmol, 33%).

Step 2: Preparation of6,8-dimethoxy-2-thioxo-2H-chromeno[2,3-d]pyrimidin-4(3H)-one

In a scintillation vial, a suspension of compound 18a, (154 mg, 0.5mmol) in a mixture of acetic acid (2.7 ml) and acetic anhydride (0.3 ml)was stirred and heated to 80° C. for 4 hours. The mixture was cooled toroom temperature. Red solid was filtered, washed with water and vacuumdried at 40° C. to 50° C. to yield compound 18 (85 mg, 0.29 mmol, 58%).

Preparation of 8-propoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step1: Preparation of 4-n-Propoxy-2-hydroxybenzaldehyde, 19a

A suspension of 2,4-dihydroxybenzaldehyde (0.76 g, 5.5 mmol),1-iodopropane (1.02 g, 6 mmol), potassium carbonate (7.60 g, 5.5 mmol),and 18-crown-6 (0.132 g, 0.5 mmol) in acetone (12 ml) was stirred andheated to 65° C. for 15 hours. The mixture was cooled to roomtemperature and quenched with HCl, aq. 1M. The mixture was extractedwith ethyl acetate (50 ml). Organic layer was washed with water (50 ml)twice and then with brine (50 ml). Organic layer was dried over MgSO4,concentrated and chromatographed to yield4-n-Propoxy-2-hydroxybenzaldehyde, 19a (0.747 g, 3.85 mmol, 77%).

Step 2: Preparation of5-(2-hydroxy-4-propoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione, 19b

In a round bottom flask, a mixture of 19a (0.66 g, 4.0 mmol), barbituricacid (0.512 g, 4.0 mmol), ethanol (15 ml) and water (20 ml) was stirredat room temperature for 15 hours. The reaction mixture was concentratedto reduce volume by 30% by evaporation. After ethanol evaporation, water(30 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield compound 19b,(0.777 g, 2.68 mmol, 67%).

Step 3: Preparation of8-propoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 19b, (145 mg, 0.5mmol) in a mixture of acetic acid (2.5 g) and acetic anhydride (0.31 g,3 mmol) was stirred and heated to 80° C. for 15 hours. The mixture wascooled to room temperature. Yellow solid was filtered, washed with waterand vacuum dried at 40° C. to 50° C. to yield compound 19 (42 mg, 0.15mmol, 30%).

Preparation of7-chloro-8-propoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 5-chloro-2,4-dihydroxybenzaldehyde

In a scintillation vial, 2,4-dihydroxybenzaldehyde (1.38 g, 10 mmol) wasdissolved in ethyl ether (18 ml). The mixture was placed under N2atmosphere and cooled to 0° C. To the mixture was added sulfurylchloride (0.91 ml, 11 mmol). The mixture was kept under N2 atmosphere at0° C. for 30 minutes. The reaction mixture was poured into ice water andextracted with ethyl acetate (20 ml). Organic layer was washed withbrine, concentrated and chromatographed to yield5-chloro-2,4-dihydroxybenzaldehyde (0.55 g, 3.2 mmol, 32%) and3-chloro-2,4-dihydroxybenzaldehyde (0.233 g, 1.34 mmol, 13.4%)

Step 2: Preparation of 5-chloro-2-hydroxy-4-propoxybenzaldehyde

A suspension of 5-chloro-2,4-dihydroxybenzaldehyde (0.517 g, 3.0 mmol),1-iodopropane (0.501 g, 3 mmol), potassium carbonate (0.414 g, 3.0mmol), and 18-crown-6 (0.079 g, 0.3 mmol) in acetone (10 ml) was stirredand heated to 70° C. for 15 hours. The mixture was cooled to roomtemperature and quenched with HCl, aq. 1M. The mixture was extractedwith ethyl acetate (30 ml). Organic layer was washed with saturatedNaHCO3 aq. (30 ml), water (30 ml) and then with brine (30 ml). Organiclayer was dried over MgSO4, concentrated and chromatographed to yield5-chloro-2-hydroxy-4-propoxybenzaldehyde (0.41 g, 1.91 mmol, 63%).

Step 3: Preparation of5-(5-chloro-2-hydroxy-4-propoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,20a

In a round bottom flask, a mixture of5-chloro-2-hydroxy-4-propoxybenzaldehyde (0.41 g, 1.91 mmol), barbituricacid (0.256 g, 2.0 mmol), ethanol (15 ml) and water (10 ml) was stirredat room temperature for 5 hours. The reaction mixture was concentratedto reduce volume by 30% by evaporation. After ethanol evaporation, water(20 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield compound 20a,(0.59 g, 1.82 mmol, 95%).

Step 4: Preparation of7-chloro-8-propoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 20a, (162 mg, 0.5mmol) in a mixture of acetic acid (2.5 g) and acetic anhydride (0.31 g,3 mmol) was stirred and heated to 80° C. for 6 hours. The mixture wascooled to room temperature. Yellow solid was filtered, washed with waterand vacuum dried at 40° C. to 50° C. to yield compound 20 (126 mg, 0.40mmol, 80%).

Preparation of 8-(n-hexyloxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1: Preparation of4-(n-hexyloxy)-2-hydroxybenzaldehyde

A suspension of 2,4-dihydroxybenzaldehyde (0.828 g, 6.0 mmol),1-iodohexane (1.40 g, 6.6 mmol), potassium carbonate (0.497 g, 3.6mmol), and 18-crown-6 (0.158 g, 0.6 mmol) in acetone (15 ml) was stirredand heated to 70° C. for 15 hours. The mixture was cooled to roomtemperature and quenched with HCl, aq. 1M. The mixture was extractedwith ethyl acetate (50 ml). Organic layer was washed with water (50 ml)twice and then with brine (50 ml). Organic layer was dried over MgSO4,concentrated and chromatographed to yield4-(n-hexyloxy)-2-hydroxybenzaldehyde (0.973 g, 4.38 mmol, 73%).

Step 2: Preparation of5-(4-(n-hexyloxy)-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,21a

In a round bottom flask, a mixture of4-(n-hexyloxy)-2-hydroxybenzaldehyde (0.444 g, 2.0 mmol), barbituricacid (0.256 g, 2.0 mmol), ethanol (15 ml) and water (15 ml) was stirredat room temperature for 3 hours. The reaction mixture was concentratedto reduce volume by 30% by evaporation. After ethanol evaporation, water(30 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield compound 21a,(0.564 g, 1.70 mmol, 85%).

Step 3: Preparation of8-(n-hexyloxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 21a, (332 mg, 1.0mmol) in a mixture of acetic acid (3.0 g) and acetic anhydride (0.51 g,5 mmol) was stirred and heated to 80° C. for 5 hours. The mixture wascooled to room temperature. Yellow solid was filtered, washed with waterand vacuum dried at 40° C. to 50° C. to yield compound 21 (148 mg, 0.46mmol, 46%).

Preparation of9-chloro-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 3-chloro-2,4-dihydroxybenzaldehyde

In a scintillation vial, 2,4-dihydroxybenzaldehyde (1.38 g, 10 mmol) wasdissolved in ethyl ether (18 ml). The mixture was placed under N2atmosphere and cooled to 0° C. To the mixture was added sulfurylchloride (0.91 ml, 11 mmol). The mixture was kept under N2 atmosphere at0° C. for 30 minutes. The reaction mixture was poured into ice water andextracted with ethyl acetate (20 ml). Organic layer was washed withbrine, concentrated and chromatographed to yield5-chloro-2,4-dihydroxybenzaldehyde (0.55 g, 3.2 mmol, 32%) and3-chloro-2,4-dihydroxybenzaldehyde (0.233 g, 1.34 mmol, 13.4%)

Step 2: Preparation of 3-chloro-2-hydroxy-4-ethoxybenzaldehyde

A suspension of 3-chloro-2,4-dihydroxybenzaldehyde (0.465 g, 2.7 mmol),iodoethane (0.421 g, 2.7 mmol), potassium carbonate (0.226 g, 1.62mmol), and 18-crown-6 (0.071 g, 0.27 mmol) in acetone (10 ml) wasstirred and heated to 70° C. for 15 hours. The mixture was cooled toroom temperature and quenched with HCl, aq. 1M. The mixture wasextracted with ethyl acetate (30 ml). Organic layer was washed withsaturated NaHCO3 aq. (30 ml), water (30 ml) and then with brine (30 ml).Organic layer was dried over MgSO4, concentrated and chromatographed toyield 3-chloro-2-hydroxy-4-ethoxybenzaldehyde (0.217 g, 1.08 mmol, 40%)and 3-chloro-2,4-diethoxybenzaldehyde (0.057 g, 0.25 mmol, 9.4%).

Step 3: Preparation of5-(3-chloro-4-ethoxy-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,22a

In a round bottom flask, a mixture of3-chloro-2-hydroxy-4-ethoxybenzaldehyde (0.217 g, 1.08 mmol), barbituricacid (0.139 g, 1.08 mmol), ethanol (10 ml) and water (10 ml) was stirredat room temperature for 10 hours. The reaction mixture was concentratedto reduce volume by 30% by evaporation. After ethanol evaporation, water(20 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake wascollected and vacuum dried at 40° C. to 50° C. to yield compound 22a,(0.30 g, 0.97 mmol, 90%).

Step 4: Preparation of9-chloro-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 22a, (155 mg, 0.5mmol) in a mixture of acetic acid (1.5 g) and acetic anhydride (0.306 g,3 mmol) was stirred and heated to 80° C. for 16 hours. The mixture wascooled to room temperature. Yellow solid was filtered, washed with waterand vacuum dried at 40° C. to 50° C. to yield compound 22 (127 mg, 0.40mmol, 80%).

Preparation of 8-isobutoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1: Preparation of 2-hydroxy-4-isobutoxybenzaldehyde

A suspension of 2,4-dihydroxybenzaldehyde (0.76 g, 5.5 mmol),1-iodo-2-methylpropane (1.196 g, 6.5 mmol), potassium carbonate (0.828g, 6.0 mmol), and 18-crown-6 (0.132 g, 0.5 mmol) in acetone (10 ml) wasstirred and heated to 70° C. for 3 days. The mixture was cooled to roomtemperature and quenched with HCl, aq. 1M. The mixture was extractedwith ethyl acetate (50 ml). Organic layer was washed with water (50 ml)twice and then with brine (50 ml). Organic layer was dried over MgSO4,concentrated and chromatographed to yield2-hydroxy-4-isobutoxybenzaldehyde (0.425 g, 2.2 mmol, 40%).

Step 2: Preparation of5-(2-hydroxy-4-isobutoxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,23a

In a round bottom flask, a mixture of 2-hydroxy-4-isobutoxybenzaldehyde(0.388 g, 2.0 mmol), barbituric acid (0.256 g, 2.0 mmol), ethanol (5 ml)and water (10 ml) was stirred at room temperature for 5 hours. Thereaction mixture was concentrated to reduce volume by 30% byevaporation. After ethanol evaporation, water (30 ml) was added to thereaction mixture. The reaction was stirred at room temperature for 30minutes and then filtered. Solid yellow cake was collected and vacuumdried at 40° C. to 50° C. to yield compound 23a, (0.563 g, 1.85 mmol,92%).

Step 3: Preparation of8-isobutoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 23a, (152 mg, 0.5mmol) in a mixture of acetic acid (1.5 g) and acetic anhydride (0.31 g,3 mmol) was stirred and heated to 80° C. for 6 hours. The mixture wascooled to room temperature. Yellow solid was filtered, andchromatographed (MeOH:DCM, 5:95) to yield compound 23 (17 mg, 0.06 mmol,12%).

Preparation of3-(n-butyl)-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 1-butylpyrimidine-2,4,6(1H,3H,5H)-trione

In a round bottom flask, dimethyl malonate (6.60 g, 50 mmol) was addedto a mixture of n-Butyl urea (5.80 g, 50 mmol) and sodium ethoxide (21%in ethanol, 19.43 g, 60 mmol). Upon the addition of dimethyl malonate,the reaction mixture was placed under nitrogen and heated to reflux.After 1 hour at reflux, the reaction mixture turned a viscoussuspension. Ethanol (15 ml) was added into the mixture. The mixture wasstirred at reflux for a total reaction time of 18 hours. At the end ofthe reaction, solid was filtered (crop 1). Filtrate was collected andconcentrated to cause precipitation. The second crop of solid wasfiltered. Combined solid crop 1 and crop 2 was dissolved in water (100ml) and acidified with HCl, aq. 1M to cause precipitation of desiredproduct. Filtrate residue from crop 2 was concentrate andchromatographed to yield desired product. Combined desired product(7.103 g, 38 mmol, 77%) was vacuum dried.

Step 2: Preparation of1-butyl-5-(4-ethoxy-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,24a

In a round bottom flask, a mixture of 2-hydroxy-4-ethoxybenzaldehyde(0.498 g, 3.0 mmol), 1-butylpyrimidine-2,4,6(1H,3H,5H)-trione (0.555 g,5.0 mmol), ethanol (10 ml) and water (10 ml) was stirred at roomtemperature for 15 hours. The reaction mixture was concentrated toreduce volume by 30% by evaporation. After ethanol evaporation, water(30 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake wascollected, washed with a mixture of ethyl acetate:hexane (20:80) andvacuum dried at 40° C. to 50° C. to yield compound 24a, (0.7939 g, 1.39mmol, 80%).

Step 3: Preparation of3-(n-butyl)-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 24a, (0.498 mg, 0.5mmol) in a mixture of acetic acid (3.5 g) and acetic anhydride (0.765 g,7.5 mmol) was stirred and heated to 80° C. for 15 hours. The mixturebecame homogeneous after 15 minutes at 80° C. The mixture was cooled toroom temperature. Yellow solid was filtered, and chromatographed(MeOH:DCM, 5:95) to yield compound 24 (102 mg, 0.32 mmol, 21%).

Preparation of7-chloro-8-propyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 4-chloro-3-propylphenol

In a scintillation vial, 3-n-Propylphenol (1.36 g, 10 mmol) wasdissolved in ethyl ether (10 ml). The mixture was cooled with ice/brinebath. To the mixture was added sulfuryl chloride (0.91 ml, 11 mmol) over2 minutes. The mixture remained at −10° C. to 0° C. for an hour. Themixture was quenched with water and extracted with ethyl acetate. Theorganic layer was concentrated and chromatographed to yield4-chloro-3-propylphenol (1.156 g, 6.8 mmol, 68%),2-chloro-3-propylphenol (0.097 g, 0.57 mmol, 5.7%) and6-chloro-3-propylphenol (0.156 g, 0.92 mmol, 9.2%).

Step 2: Preparation of 5-chloro-2-hydroxy-4-propylbenzaldehyde

In a scintillation vial, a mixture of 4-chloro-3-propylphenol (0.857 g,5 mmol), magnesium chloride anhydrous (0.712 g, 7.5 mmol),paraformaldehyde (0.90 g, 30 mmol), and triethyl amine (1.01 g, 10 mmol)in acetonitrile anhydrous (10 ml) was heated to 70° C. for 15 hours. Thereaction mixture was cooled to room temperature and then filtered. Thewhite cake was washed with HCl aq. 1M (50 ml) and ethyl acetate (80 ml).Filtrate was decanted. Organic layer was separated and washed withbrine. Organic layer was concentrate and chromatographed to yield5-chloro-2-hydroxy-4-propylbenzaldehyde (0.612 g, 3.1 mmol, 61%).

Step 3: Preparation of5-(5-chloro-2-hydroxy-4-propylbenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione

In a round bottom flask, a mixture of5-chloro-2-hydroxy-4-propylbenzaldehyde (0.397 g, 2.0 mmol), barbituricacid (0.256 g, 2.0 mmol), ethanol (10 ml) and water (10 ml) was stirredat room temperature for 15 hours. The reaction mixture was concentratedto reduce volume by 30% by evaporation. After ethanol evaporation, water(40 ml) was added to the reaction mixture. The reaction was stirred atroom temperature for 30 minutes and then filtered. Solid yellow cake waswashed with ethyl acetate:hexane (5:95), collected and vacuum dried at40° C. to 50° C. to yield compound 25a, (0.41 g, 1.32 mmol, 66%).

Step 4: Preparation of7-chloro-8-propyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 25a, (0.308 mg, 1.0mmol) in a mixture of acetic acid (2.5 g) and acetic anhydride (0.612 g,6.0 mmol) was stirred and heated to 80° C. for 6 hours. The mixture wascooled to room temperature and allowed to sit overnight. Yellow solidwas filtered, dried at 40° C. to 50° C. to yield compound 25 (250 mg,0.86 mmol, 86%).

Preparation of3-butyl-7-chloro-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1: Preparation of1-butyl-5-(5-chloro-4-ethoxy-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,26a

In a round bottom flask, a mixture of5-chloro-4-ethoxy-2-hydroxybenzaldehyde (0.295 g, 1.475 mmol),barbituric acid (0.192 g, 1.5 mmol), ethanol (20 ml) and water (10 ml)was stirred at room temperature for 3.5 hours. The reaction mixture wasconcentrated to reduce volume by 30% by evaporation. After ethanolevaporation, water (50 ml) was added to the reaction mixture. Thereaction was stirred at room temperature for 30 minutes and thenfiltered. Solid orange cake was collected and vacuum dried at 40° C. to50° C. to yield compound 26a, (0.463 g, 1.26 mmol, 85%).

Step 2: Preparation of3-butyl-7-chloro-8-ethoxy-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione,compound 26

In a scintillation vial, a suspension of compound 26a, (0.367 mg, 1.0mmol) in a mixture of acetic acid (2.5 g) and acetic anhydride (0.51 g,5.0 mmol) was stirred and heated to 80° C. for 15 hours. The mixture wascooled to room temperature and allowed to sit overnight. Yellow solidwas filtered, dried at 40° C. to 50° C. to yield compound 26 (278 mg,0.80 mmol, 80%).

Preparation of7-fluoro-8-methyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 5-fluoro-2-hydroxy-4-methylbenzaldehyde, 27a

In a scintillation vial, a suspension of 4-fluoro-3-methylphenol, (1.26g, 10.0 mmol), MgCl₂ anhydrous (1.42 g, 15 mmol), paraformaldehyde (1.50g, 50 mmol) and triethylamine (2.02 g, 20 mmol) in acetonitrileanhydrous (15 ml) was stirred and heated to 70° C. for 8 hours. Themixture was cooled to room temperature, quenched with HCl, aq. 1M (20ml) and extract with ethyl acetate. The organic layer was dried withMgSO4, filtered over silica gel, concentrated and chromatographed toyield compound 27a (1.05 g, 6.8 mmol, 68%).

Step 2: Preparation of5-(5-fluoro-2-hydroxy-4-methylbenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione

In a round bottom flask, a mixture of 27a (0.308 g, 2.0 mmol),barbituric acid (0.256 g, 2.0 mmol), ethanol (10 ml) and water (10 ml)was stirred at room temperature for 5 hours After 5 hours of reaction,water (50 ml) was added to the reaction mixture. The reaction wasstirred at room temperature for 30 minutes and then filtered. Solidyellow cake was collected and vacuum dried at 40° C. to 50° C. to yieldcompound 27b, (0.3424 g, 1.29 mmol, 65%).

Step 3: Preparation of7-fluoro-8-methyl-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 27b, (0.264 mg, 1.0mmol) in a mixture of acetic acid (4 ml) and acetic anhydride (0.51 g,5.0 mmol) was stirred and heated to 80° C. for 5 hours. The mixture wascooled to room temperature and allowed to sit overnight. Yellow solidwas filtered, dried at 40° C. to 50° C. to yield compound 27 (189 mg,0.77 mmol, 77%).

Preparation of8-ethoxy-7-fluoro-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1:Preparation of 3-ethoxy-4-fluorophenol, 28a

In a scintillation vial, a mixture of 4-fluororesorcinal (0.512 g, 4.0mmol), K2CO3 (0.607 g, 4.4 mmol), 18-crown-6 (0.106 g, 0.4 mmol),iodoethane (0.686 g, 4.4 mmol) in acetone (5 ml) was heated to 50° C.overnight. The mixture was cooled to room temperature, quenched with HClaq. 1M (20 ml), extracted with ethyl acetate (30 ml). Organic layer wasconcentrated and chromatographed to yield 28a (0.349 g, 2.34 mmol, 56%).

Step 2: Preparation of 4-ethoxy-5-fluoro-2-hydroxybenzaldehyde, 28b

In a scintillation vial, a suspension of 28a, (0.68 g, 4.3 mmol), MgCl2anhydrous (0.613 g, 6.45 mmol), paraformaldehyde (0.774 g, 25.8 mmol)and triethylamine (0.868 g, 8.6 mmol) in acetonitrile anhydrous (10 ml)was stirred and heated to 70° C. for 4 hours. The mixture was cooled toroom temperature, quenched with HCl, aq. 1M (20 ml) and extract withethyl acetate (30 ml). The organic layer was dried with MgSO4, filteredover silica gel, concentrated and chromatographed to yield compound 28b(0.70 g, 3.48 mmol, 81%).

Step 3: Preparation of5-(4-ethoxy-5-fluoro-2-hydroxybenzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,28c

In a round bottom flask, a mixture of 28b (0.184 g, 1.0 mmol),barbituric acid (0.128 g, 1.0 mmol), ethanol (10 ml) and water (10 ml)was stirred at room temperature for 6 hours After 6 hours of reaction,water (50 ml) was added to the reaction mixture. The reaction wasstirred at room temperature for 30 minutes and then filtered. Solidyellow cake was collected and vacuum dried at 40° C. to 50° C. to yieldcompound 28c, (0.263 g, 0.89 mmol, 89%).

Step 4: Preparation of8-ethoxy-7-fluoro-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 28c, (0.147 mg, 0.5mmol) in a mixture of acetic acid (2 ml) and acetic anhydride (0.204 g,2 mmol) was stirred and heated to 80° C. for 4 hours. The mixture wascooled to room temperature and allowed to sit overnight. Yellow solidwas filtered, dried at 40° C. to 50° C. to yield compound 28 (130 mg,0.48 mmol, 96%).

Synthesis of compounds 29 to 36 follow the same protocol described forcompound 28.

Synthesis of compounds 37 to 38 follow the same protocol described forcompound 20.

Synthesis of compounds 39 to 48 follow the same protocol described forcompound 22.

8-ethoxy-2H-thiochromeno[2,3-d]pyrimidine-2,4(3H)-dione Preparation of8-ethoxy-2H-thiochromeno[2,3-d]pyrimidine-2,4(3H)-dione Step 1,Preparation of2,4-dichloro-6-((3-ethoxyphenyl)thio)pyrimidine-5-carbaldehyde, 49a

In a scintillation vial, 2,4,6-trichloropyrimidine carboxaldehyde (0.422g, 2 mmol) was dissolved in anhydrous THF (5 ml). The mixture was cooledto −30° C. To the cold mixture was added triethyl amine (0.242 g, 2.4mmol) and 3-ethoxybenzenethiol (0.308 g, 2 mmol). The mixture wasmaintained at −30° C. for 3 hours. After 3 hours, the mixture wasfiltered. Cake was rinsed with ethyl ether. Filtrate contains desiredproduct was concentrated. Product was crystallized on siting undernitrogen stream. Yellow pale solid was filtered and dried under vacuumto yield 49a (0.331 g, 1.0 mmol, 50%).

Step 2, Preparation of8-ethoxy-2H-thiochromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, compound 49a (0.331 g, 1.0 mmol) was slowly andcarefully added to concentrate 98% H₂SO₄ (4 ml). The mixture was stirredat room temperature overnight. To the mixture was added EtOAc (25 ml)and water (25 ml). Precipitation occurred. Orange solid was filtered,washed with water and vacuum dried to yield compound 49 (0.137 g, 0.5mmol, 50%).

Synthesis of compounds 50-52 follow the same protocol described forcompound 22.

6-((9-chloro-2,4-dioxo-3,4-dihydro-2H-chromeno[2,3-d]pyrimidin-8-yl)oxy)hexanoicacid Step 1, Preparation of methyl6-(2-chloro-4-formyl-3-hydroxyphenoxy)hexanoate, 53a

In a scintillation vial, a suspension of3-chloro-2,4-dihydroxybenzaldehyde (0.69 g, 4 mmol), K₂CO₃ (0.552 g, 4.0mmol), 18-crown-6 (0.106 g, 0.4 mmol) and methyl 6-bromohexanoate (0.919g, 4.4 mmol) in acetone (5 ml) was heated and stirred overnight at 65°C. The reaction mixture was cooled to room temperature, acidified withHCl, aq. 1M and extracted with ethyl acetate (50 ml), twice. Organiclayer was washed with water (50 ml) twice and then with brine (50 ml).Organic layer was dried over MgSO₄, concentrated and chromatographed toyield compound 53a (0.25 g, 0.83 mmol, 20.7%).

Step 2, Preparation of 6-(2-chloro-4-formyl-3-hydroxyphenoxy)hexanoicacid, 53b

A mixture of compound 53a (0.25 g, 0.83 mmol) in MeOH (3 ml) and NaOHaq. 1M (5 ml) was warmed to 60° C. for 2 hours. The mixture was cooledto room temperature. To the mixture was added HCl, aq. 1M (8 ml) andthen extracted with ethyl acetate (20 ml). Organic layer was dried overMgSO₄, concentrated and chromatographed to yield compound 53b (0.192 g,0.67 mmol, 81%).

Step 3, Preparation of6-(2-chloro-3-hydroxy-4-((2,4,6-trioxotetrahydropyrimidin-5(2H)-ylidene)methyl)phenoxy)hexanoicacid, 53c

In a round bottom flask, a mixture of compound 53b (0.192 g, 0.67 mmol),barbituric acid (0.103 g, 0.8 mmol), iPrOH (6 ml) and water (6 ml) wasstirred at room temperature for 24 hours. Water (30 ml) was added to thereaction mixture. The reaction was stirred at room temperature for 30minutes and then filtered. Solid yellow cake was collected and vacuumdried at 40° C. to 50° C. to yield compound 53c, (0.22 g, 0.55 mmol,83%).

Step 4, Preparation of6-((9-chloro-2,4-dioxo-3,4-dihydro-2H-chromeno[2,3-d]pyrimidin-8-yl)oxy)hexanoicacid

In a scintillation vial, a suspension of compound 53c, (0.22 g, 0.55mmol) in a mixture of acetic acid (1.62 g) and acetic anhydride (0.283g) was stirred and heated to 80° C. for 2 days. The mixture was cooledto room temperature and sit for 3 hours. Yellow solid was filtered,washed with water and vacuum dried at 40° C. to 50° C. to yield compound53 (161 mg, 0.43 mmol, 78%).

Synthesis of compounds 54-59 follow the same protocol described forcompound 53.

4-((9-chloro-2,4-dioxo-3,4-dihydro-2H-chromeno[2,3-d]pyrimidin-8-yl)oxy)butylacetate Step 1, 6-(2-chloro-4-formyl-3-hydroxyphenoxy)butyl acetate, 60a

In scintillation vial, a suspension of3-chloro-2,4-dihydroxybenzaldehyde (0.5175 g, 3 mmol), KHCO₃ (0.49 g,4.9 mmol), NaI (0.45 g, 3 mmol), 18-crown-6 (0.132 g, 0.5 mmol) andmethyl 6-bromobutanoate (0.585 g, 3.0 mmol) in acetone (5 ml) was heatedand stirred for 4 days at 70° C. The reaction mixture was cooled to roomtemperature, acidified with HCl, aq. 1M and extracted with ethyl acetate(25 ml). Organic layer was washed with water (25 ml) and then with brine(25 ml). Organic layer was dried over MgSO₄, concentrated andchromatographed to yield compound 60a (0.182 g, 0.64 mmol, 21.2%).

Step 2, Preparation of4-(2-chloro-3-hydroxy-4-((2,4,6-trioxotetrahydropyrimidin-5(2H)-ylidene)methyl)phenoxy)butylacetate, 60b

In a scintillation vial, a mixture of compound 60a (0.182 g, 0.64 mmol),barbituric acid (0.089 g, 0.7 mmol), iPrOH (3 ml) and water (3 ml) wasstirred at room temperature for 24 hours. Water (10 ml) was added to thereaction mixture. The reaction was stirred at room temperature for 10minutes and then filtered. Solid yellow cake was collected and vacuumdried at 40° C. to 50° C. to yield compound 60b, (0.209 g, 0.53 mmol,82%).

Step 3, Preparation of4-((9-chloro-2,4-dioxo-3,4-dihydro-2H-chromeno[2,3-d]pyrimidin-8-yl)oxy)butylacetate

In a scintillation vial, a suspension of compound 60b, (0.209 g, 0.55mmol) in a mixture of acetic acid (1.65 g) and acetic anhydride (0.281g) was stirred and heated to 80° C. for 4.5 hours. The mixture wascooled to room temperature and sit for 3 hours. Yellow solid wasfiltered, dried at 40° C. to 50° C. to yield compound 60 (108 mg, 0.28mmol, 52%).

8-(2-(2-methoxyethoxy)ethoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1, Preparation of2-hydroxy-4-(2-(2-methoxyethoxy)ethoxy)benzaldehyde, 61a

In scintillation vial, a suspension of 2,4-dihydroxybenzaldehyde (0.552g, 4 mmol), K₂CO₃ (0.552 g, 4.0 mmol), 18-crown-6 (0.106 g, 0.4 mmol)and 1-bromo-2-(2-methoxyethoxy) ethane (90-95% purity, 0.61 g, 3.0 mmol)in acetone (5 ml) was heated and stirred overnight at 80° C. Thereaction mixture was cooled to room temperature, acidified with HCl, aq.1M and extracted with ethyl acetate (25 ml). Organic layer was washedwith water (25 ml) and then with brine (25 ml). Organic layer was driedover MgSO₄, concentrated and chromatographed to yield compound 61a (0.35g, 1.46 mmol, 36.4%).

Step 2, Preparation of5-(2-hydroxy-4-(2-(2-methoxyethoxy)ethoxy)benzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,61b

In a scintillation vial, a mixture of compound 61a (0.35 g, 1.46 mmol),barbituric acid (0.224 g, 1.75 mmol), ethanol (10 ml) and water (10 ml)was stirred and warmed to 40 to 50° C. for 15 minutes and them stirredat room temperature overnight. Water (30 ml) was added to the reactionmixture. The reaction was stirred at room temperature for 15 minutes andthen filtered. Solid yellow cake was collected and vacuum dried at 40°C. to 50° C. to yield compound 61b, (0.434 g, 1.24 mmol, 85%).

Step 3, Preparation of8-(2-(2-methoxyethoxy)ethoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 61b, (0.263 g, 0.75mmol) in a mixture of acetic acid (2.25 g) and acetic anhydride (0.383g) was stirred and heated to 80° C. for 6 hours. The mixture wasconcentrated to remove ˜50% of its volume and then cooled to roomtemperature and sit for 3 hours. Yellow solid was filtered, dried at 40°C. to 50° C. to yield compound 61 (189 mg, 0.57 mmol, 76%).

Synthesis of compounds of example 62-65 follow the same protocoldescribed for compound 61.

9-chloro-8-(4-methoxybutoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1, Preparation of3-chloro-2-hydroxy-4-(4-methoxybutoxy)benzaldehyde, 66a

In scintillation vial, a suspension of3-chloro-2,4-dihydroxybenzaldehyde (1.725 g, 10 mmol), K₂CO₃ (1.38 g, 10mmol), 18-crown-6 (0.264 g, 1.0 mmol) and 1-bromo-4-methoxybutane (0.835g, 5.0 mmol) in acetone (10 ml) was heated and stirred overnight at 70°C. The reaction mixture was cooled to room temperature, acidified withHCl, aq. 1M and extracted with ethyl acetate (50 ml). Organic layer waswashed with water (50 ml) and then with brine (50 ml). Organic layer wasdried over MgSO₄, concentrated and chromatographed to yield compound 66a(0.898 g, 3.47 mmol, 34.7%).

Step 2, Preparation of5-(3-chloro-2-hydroxy-4-(4-methoxybutoxy)benzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,66b

In a round bottom flash, a mixture of compound 66a (0.498 g, 1.5 mmol),barbituric acid (0.256 g, 2.0 mmol), ethanol (6 ml) and water (6 ml) wasstirred at room temperature overnight. Water (15 ml) was added to thereaction mixture. The reaction was stirred at room temperature for 15minutes and then filtered. Solid yellow cake was collected and vacuumdried at 40° C. to 50° C. to yield compound 66b, (0.466 g, 1.34 mmol,89.6%).

Step 3, Preparation of9-chloro-8-(4-methoxybutoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 66b, (0.369 g, 1.0mmol) in a mixture of acetic acid (2.1 g) and acetic anhydride (0.51 g)was stirred and heated to 80 to 90° C. overnight. The mixture wasconcentrated to remove ˜50% of its volume and then cooled to roomtemperature and sit for 3 hours. Yellow solid was filtered, dried at 40°C. to 50° C. to yield compound 66 (300 mg, 0.86 mmol, 86%).

Synthesis of compounds of example 67-70 follow the same protocoldescribed for compound 66.

Synthesis of compounds 71-74 follow the same protocol described forcompound 75, below.

7-fluoro-8-(3-(2-methoxyethoxy)propoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dioneStep 1, Preparation of 4-fluoro-3-(3-(2-methoxyethoxy)propoxy)phenol,75a

In scintillation vial, a suspension of 4-fluororesorcinol (1.28 g, 10mmol), K₂CO₃ (1.38 g, 10 mmol), 18-crown-6 (0.264 g, 1.0 mmol) and1-bromo-3-(2-methoxyethoxy)propane (0.985 g, 5.0 mmol) in acetone (10ml) was heated and stirred overnight at 70° C. The reaction mixture wascooled to room temperature, acidified with HCl, aq. 1M and extractedwith ethyl acetate (50 ml). Organic layer was washed with water (50 ml)and then with brine (50 ml). Organic layer was dried over MgSO₄,concentrated and chromatographed to yield compound 75a (0.921 g, 3.74mmol, 37.4%).

Step 2, Preparation of5-fluoro-2-hydroxy-4-(3-(2-methoxyethoxy)propoxy)benzaldehyde, 75b

In a scintillation vial, a suspension of compound 75a, (0.478 g, 1.96mmol), MgCl₂ anhydrous (0.279 g, 2.93 mmol), paraformaldehyde (0.12 g, 4mmol) and triethylamine (0.296 g, 2.93 mmol) in acetonitrile anhydrous(5 ml) was stirred and heated to 60° C. for 2 hours. The mixture wascooled to room temperature, quenched with HCl, aq. 1M (15 ml) andextract with ethyl acetate. The organic layer was dried with MgSO₄,filtered over silica gel, concentrated and chromatographed to yieldcompound 75b (0.34 g, 1.25 mmol, 64%).

Step 3, Preparation of5-(5-fluoro-2-hydroxy-4-(3-(2-methoxyethoxy)propoxy)benzylidene)pyrimidine-2,4,6(1H,3H,5H)-trione,75c

In a round bottom flash, a mixture of compound 75b (0.140 g, 0.51 mmol),barbituric acid (0.079 g, 0.62 mmol), ethanol (3 ml) and water (3 ml)was stirred at room temperature overnight. Water (15 ml) was added tothe reaction mixture. The reaction was stirred at room temperature for15 minutes and then filtered. Solid yellow cake was collected and vacuumdried at 40° C. to 50° C. to yield compound 75c, (0.189 g, 0.49 mmol,97%).

Step 4, Preparation of7-fluoro-8-(3-(2-methoxyethoxy)propoxy)-2H-chromeno[2,3-d]pyrimidine-2,4(3H)-dione

In a scintillation vial, a suspension of compound 75c, (0.172 g, 0.45mmol) in a mixture of acetic acid (1.58 g) and acetic anhydride (0.226g) was stirred and heated to 80 to 90° C. for 5 hours. The mixture wasconcentrated to remove ˜50% of its volume and then cooled to roomtemperature and sit for 3 hours. Yellow solid was filtered, washed withwater and vacuum dried at 40° C. to 50° C. to yield compound 75 (144 mg,0.39 mmol, 88%).

NMR Spectroscopy

NMR spectroscopy was performed using standard Bruker® 400 MHz NMR.

Cmpd. Structure MW 1H NMR, 400 MHz, DMSO-d6 1

244 3.99 (s, 3H), 7.48 (dd, J = 7.92, 7.92 Hz, 1H), 7.58-7.63 (m, 2H),8.92 (s, 1H), 11.46 (s, 1H) 2

258 1.44 (t, J = 7.00 Hz, 3H), 4.26 (q, J = 7.00 Hz, 2H), 7.46 (dd, J =7.92, 7.92 Hz, 1H), 7.57-7.63 (m, 2H), 8.92 (s, 1H), 11.47 (s, 1H) 3

279 4.02 (s, 3H), 7.65 (d, J = 2.30 Hz, 1H), 7.73 (d, J = 2.30 Hz, 1H),8.85 (s, 1H), 11.53 (s, 1H) 4

323 4.01 (s, 3H), 7.73 (d, J = 2.14 Hz, 1H), 7.87 (d, J = 2.14 Hz, 1H),8.83 (s, 1H), 11.52 (s, 1H) 5

293 1.43 (t, J = 7.00 Hz, 3H), 4.28 (q, J = 7.00 Hz, 2H), 7.63 (d, J =2.28 Hz, 1H), 7.72 (d, J = 2.28 Hz, 1H), 8.84 (s, 1H), 11.53 (s, 1H) 6

244 3.91 (s, 3H), 7.11 (dd, J = 8.88, 2.65 Hz, 1H), 7.28 (d, J = 2.65Hz, 1H), 7.95 (d, J = 8.88 Hz, 1H), 8.83 (s, 1H), 11.28 (s, 1H) 7

295 4.00 (s, 3H), 7.65 (d, J = 2.30 Hz, 1H), 7.73 (d, J = 2.30 Hz, 1H),9.01 (s, 1H), 12.63 (s, 1H) 8

260 4.00 (s, 3H), 7.24 (dd, J = 8.86, 2.43 Hz, 1H), 7.88 (d, J = 2.43Hz, 1H), 8.06 (d, J = 8.86 Hz, 1H), 9.01 (s, 1H), 12.62 (s, 1H) 9

214 7.56 (ddd, J = 8.30, 7.60, 1.00 Hz, 1H), 7.71 (d, J = 8.30 Hz, 1H),7.90 (ddd, J = 8.40, 7.30, 1.50 Hz, 1H), 8.09 (dd, J = 8.80, 1.50 Hz,1H), 8.95 (s, 1H), 11.46 (s, 1H) 10

244 3.86 (s, 3H), 7.50 (dd, J = 9.12, 3.08 Hz, 1H), 7.65 (d, J = 3.08Hz, 1H), 7.68 (d, J = 8.00 Hz, 1H), 8.88 (s, 1H), 11.43 (s, 1H) 11

230 6.98 (d, J = 2.04 Hz, 1H), 7.00 (dd, J = 8.55, 2.04 Hz, 1H), 7.93(d, J = 8.60 Hz, 1H), 8.84 (s, 1H), 11.27 (s, 1H), 11.62 (bs, 1H) 12

228 2.60 (s, 3H), 7.49 (dd, J = 8.00, 0.85 Hz, 1H), 7.65 (d, J = 0.85Hz, 1H), 8.05 (d, J = 8.00 Hz, 1H), 9.00 (s, 1H), 11.49 (s, 1H) 13

258 1.32 (t, J = 6.94 Hz, 3H), 4.19 (q, J = 6.94 Hz, 2H), 7.09 (dd, J =8.78, 2.50 Hz, 1H), 7.25 (d, J = 2.50 Hz, 1H), 7.93 (d, J = 8.78 Hz,1H), 8.81 (s, 1H), 11.26 (s, 1H) 14

274 3.98 (s, 3H), 4.01 (s, 3H), 6.71 (d, J = 1.67 Hz, 1H), 6.99 (d, J =1.67 Hz, 1H), 8.61 (s, 1H), 11.30 (s, 1H) 15

260 3.87 (s, 3H), 7.59 (dd, J = 9.13, 3.02 Hz, 1H), 7.68 (d, J = 3.02Hz, 1H), 7.73 (d, J = 9.13 Hz, 1H), 8.98 (s, 1H), 12.70 (s, 1H) 16

246 7.22 (d, J = 2.00 Hz, 1H), 7.27 (dd, J = 8.70, 2.10 Hz, 1H), 8.19(d, J = 8.80 Hz, 1H), 9.17 (s, 1H), 12.11 (bs, 1H), 12.77 (s, 1H) 17

244 2.45 (s, 3H), 7.37 (dd, J = 8.17, 1.00 Hz, 1H), 7.53 (d, J = 1.00Hz, 1H), 7.92 (d, J = 8.17 Hz, 1H), 8.94(s, 1H), 12.61 (s, 1H) 18

290 3.94 (s, 3H), 3.95 (s, 3H), 6.68 (d, J = 2.01 Hz, 1H), 6.90 (d, J =2.01 Hz, 1H), 8.60 (s, 1H), 12.53 (s, 1H) 19

272 0.81 (t, J = 7.42 Hz, 3H), 1.55-1.63 (m, 2H), 3.97 (t, J = 6.55 Hz,2H), 6.97 (dd, J = 8.81, 2.40 Hz, 1H), 7.13 (d, J = 2.40 Hz, 1H), 7.81(d, J = 8.81 Hz, 1H), 8.69 (s, 1H), 11.14 (s, 1H) 20

307 0.96 (t, J = 7.32 Hz, 3H), 1.71-1.80 (m, 2H), 4.19 (t, J = 6.38 Hz,2H), 7.50 (s, 1H), 8.16 (s, 1H), 8.76 (s, 1H), 11.33 (s, 1H) 21

314 0.87-0.91 (m, 3H), 1.31-1.35 (m, 4H), 1.40-1.45 (m, 2H), 1.74-1.79(m, 2H), 4.20 (t, J = 6.50 Hz, 2H), 7.16 (dd, J = 8.88, 2.40 Hz, 1H),7.33 (d, J = 2.40 Hz, 1H), 8.00 (d, J = 8.88 Hz, 1H), 8.89 (s, 1H),11.33 (s, 1H) 22

293 1.43 (t, J = 6.96 Hz, 3H), 4.37 (q, J = 6.96 Hz, 2H), 7.44 (d, J =8.96 Hz, 1H), 8.05 (d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.44 (s, 1H) 23

286 1.12 (s, 3H), 1.14 (s, 3H), 2.15-2.25 (m, 1H), 4.11 (d, J = 6.56 Hz,2H), 7.29 (dd, J = 8.76, 2.32, Hz, 1H), 7.45 (d, J = 2.24 Hz, 1H), 8.12(d, J = 8.84 Hz, 1H), 9.01 (s, 1H), 11.45 (s, 1H) 24

314 0.92 (t, J = 7.24 Hz, 3H), 1.26-1.33 (m, 2H), 1.40 (t, J = 6.96 Hz,3H), 1.50-1.57 (m, 2H), 3.84 (t, J = 7.32 Hz, 2H), 4.25 (q, J = 7.00 Hz,2H), 7.17 (dd, J = 8.76, 2.32, Hz, 1H), 7.34 (d, J = 2.32 Hz, 1H), 8.04(d, J = 8.84 Hz, 1H), 8.95 (s, 1H) 25

291 1.05 (t, J = 7.25 Hz, 3H), 1.64-1.70 (m, 2H), 2.81-2.85 (m, 2H),7.76 (s, 1H), 8.20 (s, 1H), 8.85 (s, 1H), 11.50 (s, 1H) 26

349 0.86 (t, J = 7.28 Hz, 3H), 1.19-1.28 (m, 2H), 1.36 (t, J = 6.92 Hz,3H), 1.43-1.50 (m, 2H), 3.76 (q, J = 7.40 Hz, 2H), 4.28 (q, J = 7.00 Hz,2H), 7.51 (s, 1H), 8.19 (s, 1H), 8.81 (s, 1H) 27

246 2.43 (d, J = 1.5 Hz, 3H), 7.75 (d, J = 6.17 Hz, 1H), 7.90 (d, J =9.17 HZ, 1H), 8.87 (s, 1H), 11.47 (s, 47, 1H) 28

276 1.42 (t, J = 6.80 Hz, 3H), 4.28 (q, J = 6.80 Hz, 2H), 7.62 (d, J =7.05 Hz, 1H), 7.97 (d, J = 10.86 Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 29

304 0.96 (t, J = 7.36 Hz, 3H), 1.43-1.49 (m, 2H), 1.76-1.83 (m, 2H),4.28 (t, J = 6.44 Hz, 2H), 7.63 (d, J = 7.04 Hz, 1H), 7.98 (d, J = 10.85Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 30

318 0.93 (t, J = 7.36 Hz, 3H), 1.34-1.44 (m, 4H), 1.77-1.84 (m, 2H),4.28 (t, J = 6.44 Hz, 2H), 7.62 (d, J = 7.04 Hz, 1H), 7.98 (d, J = 10.85Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 31

332 0.87 (t, J = 6.85 Hz, 3H), 1.30-1.35 (m, 4H), 1.39-1.48 (m, 2H),1.76- 1.82 (m, 2H), 4.28 (t, J = 6.50 Hz, 2H), 7.62 (d, J = 7.00 Hz,1H), 7.98 (d, J = 10.85 Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 32

346 0.86 (t, J = 6.80 Hz, 3H), 1.28-1.30 (m, 4H), 1.32-1.37 (m, 2H),1.42- 1.46 (m, 2H), 1.76-1.83 (m, 2H), 4.27 (t, J = 6.40 Hz, 2H), 7.62(d, J = 7.00 Hz, 1H), 7.98 (d, J = 10.80 Hz, 1H), 8.83 (s, 1H), 11.38(s, 1H) 33

318 0.95 (s, 3H), 0.97 (s, 3H), 1.68-1.73 (m, 2H), 1.77-1.82 (m, 1H),4.31 (t, J = 6.60 Hz, 2H), 7.66 (d, J = 7.04 Hz, 1H), 7.97 (d, J = 10.85Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 34

332 0.99 (s, 9H), 1.75 (t, J = 7.16 2H), 4.33 (t, J = 7.16 Hz, 2H), 7.71(d, J = 7.04 Hz, 1H), 7.98 (d, J = 10.85 Hz, 1H), 8.83 (s, 1H), 11.38(s, 1H) 35

332 0.88 (s, 3H), 0.90 (s, 3H), 1.29-1.35 (m, 2H), 1.57-1.64 (m, 1H),1.76- 1.84 (m, 2H), 4.27 (t, J = 6.60 Hz, 2H), 7.61 (d, J = 7.00 Hz,1H), 7.97 (d, J = 10.85 Hz, 1H), 8.83 (s, 1H), 11.38 (s, 1H) 36

318 1.04 (s, 9H), 3.97 (s, 2H), 7.63 (d, J = 7.40 Hz, 1H), 7.99 (d, J =10.85 Hz, 1H), 8.84 (s, 1H), 11.38 (s, 1H) 37

349 0.83 (t, J = 7.36 Hz, 3H), 1.22.-1.29 (m, 4H), 1.37-1.41 (m, 2H),4.22 (t, J = 6.40 Hz, 2H), 7.50 (s, 1H), 8.15 (s, 1H), 8.75 (s, 1H),11.32 (s, 1H) 38

321 1.02 (s, 3H), 1.04 (s, 3H), 2.09-2.15 (m, 1H), 4.08 (d, J = 6.48 Hz,2H), 7.56 (s, 1H), 8.23 (s, 1H), 8.83 (s, 1H), 11.40 (s, 1H) 39

335 0.91 (t, J = 7.00 Hz, 3H), 1.36-1.47 (m, 4H), 1.77-1.85 (m, 2H),4.30 (t, J = 6.50 Hz, 2H), 7.45 (d, J = 9.00 Hz, 1H), 8.06 (d, J = 9.00Hz, 1H), 8.88 (s, 1H), 11.43 (s, 1H) 40

335 0.93 (s, 3H), 0.96 (s, 3H), 1.69-1.74 (m, 2H), 1.81-1.88 (m, 1H),4.33 (t, J = 6.52 Hz, 2H), 7.48 (d, J = 8.96 Hz, 1H), 8.06 (d, J = 8.96Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H) 41

349 1.06 (s, 9H), 1.82 (t, J = 6.90 Hz, 2H), 4.42 (t, J = 6.90 Hz, 2H),7.59 (d, J = 9.00 Hz, 1H), 8.13 (d, J = 9.00 Hz, 1H), 8.94 (s, 1H),11.49 (s, 1H) 42

363 0.86 (t, J = 6.80 Hz, 3H), 1.27-1.30 (m, 4H), 1.32-1.37 (m, 2H),1.42- 1.46 (m, 2H), 1.78-1.83 (m, 2H), 4.30 (t, J = 6.40 Hz, 2H), 7.45(d, J = 8.96 Hz, 1H), 8.05 (d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.43 (s,1H) 43

321 0.96 (t, J = 7.5 Hz, 3H), 1.47-1.52 (m, 2H), 1.76-1.83 (m, 2H), 4.32(t, J = 6.40 Hz, 2H), 7.46 (d, J = 8.96 Hz, 1H), 8.06 (d, J = 8.96 Hz,1H), 8.89 (s, 1H), 11.43 (s, 1H) 44

349 0.88 (t, J = 7.01 Hz, 3H), 1.29-1.36 (m, 4H), 1.45-1.49 (m, 2H),1.77- 1.83 (m, 2H), 4.30 (t, J = 6.40 Hz, 2H), 7.45 (d, J = 8.96 Hz,1H), 8.05 (d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H) 45

304 1.01 (t, J = 6.80, 3H), 1.47-1.57 (m, 2H), 1.83-1.88 (m, 2H), 4.35(d, t = 6.50 Hz, 2H), 7.52 (dd, J = 7.2 Hz, 8.8 Hz, 1H), 7.96 (dd, J =1.8 hz, 9.0 Hz), 8.96 (d, J = 1.40 Hz, 1H), 11.51 (s, 1H) 46

332 0.87 (t, J = 6.80 Hz, 3H), 1.30-1.35 (m, 4H), 1.39-1.48 (m, 2H),1.76- 1.83 (m, 2H), 4.28 (t, J = 6.50 Hz, 2H), 7.46 (dd, J = 7.2 Hz, 8.8Hz, 1H), 7.90 (dd, J = 1.8 hz, 9.0 Hz), 8.89 (d, J = 1.40 Hz, 1H), 11.45(s, 1H) 47

332 0.88 (s, 3H), 0.90 (s, 3H), 1.30-1.36 (m, 2H), 1.56-1.63 (m, 1H),1.76- 1.84 (m, 2H), 4.28 (t, J = 6.60 Hz, 2H), 7.46 (dd, J = 7.2 Hz, 8.8Hz, 1H), 7.90 (dd, J = 1.8 hz, 9.0 Hz), 8.89 (d, J = 1.40 Hz, 1H), 11.45(s, 1H) 48

304 1.01 (s, 3H), 1.03(s, 3H), 2.07-2.17 (m, 1H), 4.07(d, J = 6.60 Hz,2H), 7.46 (dd, J = 7.2 Hz, 8.8 Hz, 1H), 7.90 (dd, J = 1.8 hz, 9.0 Hz),8.90 (d, J = 1.54 Hz, 1H), 11.45 (s, 1H) 49

274 1.39 (t, J = 6.96 Hz, 3H), 4.25 (q, J = 6.97 Hz, 2H), 7.25 (dd, J =8.84 Hz, 2.40 Hz, 1H), 7.63 (d, J = 2.40 Hz, 1H0, 8.24 (d, J = 8.84 Hz,1H), 8.82 (s, 1H), 11. 33 (s, 1H) 50

286 0.91 (t, J = 7.42 Hz, 3H), 1.46-1.53 (m, 2H), 1.72-1.79 (m, 2H),4.20 (t, J = 6.48 Hz, 2H), 7.16 (dd, J = 8.80 Hz, 2.20 Hz, 1H), 7.33 (d,J = 2.20 Hz, 1H), 7.99 (d, J = 8.84 Hz, 1H), 8.88 (s, 1H), 11.32 (s, 1H)51

344 1.73-1.77 (m, 2H), 1.78-1.85 (m, 2H), 2.01 (s, 3H), 4.06 (t, J =6.40 Hz, 2H) 4.23 (t, J = 6.08 Hz, 2H), 7.17 (dd, J = 8.84 Hz, 2.20 Hz,1H), 7.33 (d, J = 2.20 Hz, 1H), 8.00 (d, J = 8.84 Hz, 1H), 8.89 (s, 1H),11.32 (s, 1H) 52

346.35 0.88 (t, J = 6.40 Hz, 3H), 1.26-1.38 (m, 6H), 1.40-1.47 (m, 2H),1.77- 1.84 (m, 2H), 4.29 (t, J = 6.30 Hz, 2H), 7.46 (dd, J = 7.2 Hz, 8.8Hz, 1H), 7.89 (dd, J = 1.8 hz, 9.0 Hz), 8.90 (d, J = 1.40 Hz, 1H), 11.45(s, 1H) 53

378.76 1.46-1.51 (m, 2H), 1.56-1.63 (m, 2H), 1.79-1.91 (m, 2H),2.22-2.27 (m, 2H), 4.31 (t, J = 6.28 Hz, 2H), 7.45 (d, J = 8.96 Hz, 1H),8.05 (d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H), 12.02 (bs, 1H)54

364.74 1.66-1.74 (m, 2H), 1.80-1.86 (m, 2H), 2.33(t, J = 7.40 Hz, 2H),4.32 (t, J = 6.16 Hz, 2H), 7.45 (d, J = 8.96 Hz, 1H), 8.05 (d, J = 8.96Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H), 12.07 (bs, 1H) 55

350.71 2.01-2.08 (m, 2H), 2.46(t, J = 7.26 Hz, 2H), 4.33 (t, J = 6.26Hz, 2H), 7.45 (d, J = 8.91 Hz, 1H), 8.05 (d, J = 8.91 Hz, 1H), 8.89 (s,1H), 11.44 (s, 1H), 12.22 (bs, 1H) 56

322.66 5.10 (s, 2H), 7.36 (d, J = 9.00 Hz, 1H), 8.02 (d, J = 9.00 Hz,1H), 8.87 (s, 1H), 11.45 (s, 1H), 12.02 (bs, 1H) 57

378.76 1.70-1.77 (m, 2H), 1.81-1.87 (m, 2H), 2.43 (t, J = 7.40 Hz, 2H),3.60 (s, 3H), 4.32 (t, J = 6.10 Hz, 2H), 7.44 (d, J = 8.96 Hz, 1H), 8.05(d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H) 58

406.82 1.36-1.43 (m, 2H), 1.45-1.1.52 (m, 2H), 1.57-1.62 (m, 2H),1.78-1.85 (m, 2H), 2.00 (s, 3H), 4.00 (t, J = 6.62 6.62 Hz, 2H), 4.31(t, J = 6.30 Hz, 2H), 7.45 (d, J = 8.96 Hz, 1H), 8.05 (d, J = 8.96 Hz,1H), 8.89 (s, 1H), 11.43 (s, 1H) 59

364.74 2.02 (s, 3H), 2.11-2.18 (m, 2H), 4.21 (t, J = 6.38 Hz, 2H), 4.38(t, J = 6.10 Hz, 2H), 7.46 (d, J = 8.96 Hz, 1H), 8.06 (d, J = 8.96 Hz,1H), 8.89 (s, 1H), 11.44 (s, 1H) 60

378.76 1.75-1.82 (m, 2H), 1.84-1.89 (m, 2H), 2.01 (s, 3H), 4.09 (t, J =6.45 Hz, 2H), 4.31 (t, J = 6.04 Hz, 2H), 7.45 (d, J = 8.96 Hz, 1H), 8.06(d, J = 8.96 Hz, 1H), 8.89 (s, 1H), 11.44 (s, 1H) 61

332.31 3.25 (s, 3H), 3.44-3.48 (m, 2H), 3.58-3.62 (m, 2H), 3.78-3.81 (m,2H), 4.32-4.36 (m 2H), 7.18 (dd, J = 8.72, 2.52 Hz, 1H), 7.35 (d, J =2.52 Hz, 1H), 8.00 (d, J = 8.84 Hz, 1H), 8.88 (s, 1H), 11.33 (s, 1H) 62

366.75 3.25 (s, 3H), 3.47 (t, J = 4.72 Hz, 2H), 3.65 (t, J = 4.72 Hz,2H), 3.85 (t, J = 4.33 Hz, 2H), 4.44 (t, J = 4.33 Hz, 2H), 7.46 (d, J =8.82 Hz, 1H), 8.05 (d, J = 8.82 Hz, 1H), 8.89 (s, 1H), 11.44 (s, 1H) 63

350.3 3.24 (s, 3H), 3.47 (t, J = 4.72 Hz, 2H), 3.65 (t, J = 4.70 Hz,2H), 3.86 (t, J = 4.36 Hz, 2H), 4.42 (t, J = 4.36 Hz, 2H), 7.46 (dd, J =7.2 Hz, 8.90 Hz, 1H), 7.89 (dd, J = 1.8 hz, 9.0 Hz), 8.90 (d, J = 1.40Hz, 1H), 11.45 (s, 1H) 64

366.75 3.25 (s, 3H), 3.49 (t, J = 4.72 Hz, 2H), 3.66 (t, J = 4.74 Hz,2H), 3.86 (t, J = 4.40 Hz, 2H), 4.45 (t, J = 4.33 Hz, 2H), 7.53 (s, 1H),8.17 (s, 1H), 8.88 (s, 1H), 11.43 (s, 1H) 65

350.3 3.24 (s, 3H), 3.46 (t, J = 4.72 Hz, 2H), 3.65 (t, J = 4.66 Hz,2H), 3.86 (t, J = 4.36 Hz, 2H), 4.44 (t, J = 4.36 Hz, 2H),7.62 (d, J =7.00 Hz, 1H), 7.98 (d, J = 10.80 Hz, 1H), 8.90 (s, 1H), 11.40 (s, 1H) 66

350.75 1.67-1.73 (m, 2H), 1.82-1.89 (m, 2H), 3.25 (s, 3H), 3.40 (t, J =6.30 Hz, 2H), 4.33 (t, J = 6.30 Hz, 2H), 7.45 (d, J = 8.95 Hz, 1H), 8.08(d, J = 8.95 Hz, 1H), 8.89 (s, 1H), 11.43 (s, 1H) 67

316.31 1.65-1.70 (m, 2H), 1.77-1.83 (m, 2H), 3.25 (s, 3H), 3.38 (t, J =6.37 Hz, 2H), 4.22 (t, J = 6.37 Hz, 2H), 7.16 (dd, J = 8.72, 2.26 Hz,1H), 7.32 (d, J = 2.26 Hz, 1H), 8.00 (d, J = 8.72 Hz, 1H), 8.88 (s, 1H),11.32 (s, 1H) 68

334.3 1.66-1.71 (m, 2H), 1.81-1.89 (m, 2H), 3.24 (s, 3H), 3.40 (t, J =6.36 Hz, 2H), 4.34 (t, J = 6.36 Hz, 2H), 7.45 (dd, J = 7.2 Hz, 8.8 Hz,1H), 7.89 (dd, J = 1.8 hz, 9.0 Hz), 8.89 (d, J = 1.40 Hz, 1H), 11.45 (s,1H) 69

350.75 1.67-1.74 (m, 2H), 1.82-1.88 (m, 2H), 3.25 (s, 3H), 3.42 (t, J =6.30 Hz, 2H), 4.32 (t, J = 6.30 Hz, 2H), 7.50 (s, 1H), 8.15 (s, 1H),8.86 (s, 1H), 11.43 (s, 1H) 70

334.3 1.67-1.75 (m, 2H), 1.82-1.88 (m, 2H), 3.24 (s, 3H), 3.43 (t, J =6.33 Hz, 2H), 4.32 (t, J = 6.30 Hz, 2H), 7.65 (d, J = 7.04 Hz, 1H), 7.98(d, J = 10.86 Hz, 1H), 8.84 (s, 1H), 11.40 (s, 1H) 71

346.33 2.02-2.08 (m, 2H), 3.23 (s, 3H), 3.43-3.45 (m, 2H), 3.50-3.56 (m,2H), 3.60 (t, J = 6.20 Hz, 2H), 4.36 (t, J = 6.20 Hz, 2H), 7.17 (dd, J =8.72, 2.24 Hz, 1H), 7.33 (d, J = 2.24 Hz, 1H), 8.00 (d, J = 8.72 Hz,1H), 8.89 (s, 1H), 11.34 (s, 1H) 72

380.78 2.02-2.09 (m, 2H), 3.22 (s, 3H), 3.43-3.45 (m, 2H), 3.50-3.54 (m,2H), 3.60 (t, J = 6.12 Hz, 2H), 4.36 (t, J = 6.12 Hz, 2H), 7.46 (d, J =8.92 Hz, 1H), 8.05 (d, J = 8.82 Hz, 1H), 8.89 (s, 1H), 11.44 (s, 1H) 73

364.33 2.03-2.10 (m, 2H), 3.24 (s, 3H), 3.41-3.45 (m, 2H), 3.50-3.54 (m,2H), 3.60 (t, J = 6.18 Hz, 2H), 4.38 (t, J = 6.18 Hz, 2H),7.45 (dd, J =7.2 Hz, 8.8 Hz, 1H), 7.88 (dd, J = 1.8 hz, 9.0 Hz), 8.89 (d, J = 1.40Hz, 1H), 11.42 (s, 1H) 74

380.78 2.03-2.09 (m, 2H), 3.23 (s, 3H), 3.43-3.48 (m, 2H), 3.50-3.56 (m,2H), 3.60 (t, J = 6.18 Hz, 2H), 4.34 (t, J = 6.18 Hz, 2H), 7.52 (s, 1H),8.18(s, 1H), 8.86 (s, 1H), 11.35 (s, 1H) 75

364.33 2.00-2.08 (m, 2H), 3.24 (s, 3H), 3.43-3.46 (m, 2H), 3.51-3.54 (m,2H), 3.57 (t, J = 6.25 Hz, 2H), 4.34 (t, J = 6.12 Hz, 2H), 7.64 (d, J =7.00 Hz, 1H), 7.98 (d, J = 10.71 Hz, 1H), 8.84 (s, 1H), 11.39 (s, 1H)

Example 3 Materials and Methods

Fluorescence Polarization (FP) Assay

To identify direct NF-κB inhibitors, a fluorescence polarization (FP)screening assay was developed using c-Rel homodimer and CD28 responseelement (CD28RE) in the promoter region of the IL-2 gene.5′-fluorescein-labeled duplex CD28RE oligonucleotide probe (10 nM) wasmixed with Rel protein (128 nM) in reaction buffer (20 mM Tris (pH7.5),100 mM NaCl, 0.5 ug/ml polydldC, 1% NP-40, 0.1% BSA). 20 μl of themixture was added to each well of a 384-well plate, compound of interestwas added (25 μM), and plates were incubated for 30 minutes at roomtemperature. The anisotropy value of each reaction well was measuredusing Fusion™ Universal Microplate Analyzer (Perkin Elmer, PE). A seriesof titration experiments were performed to optimize c-Rel protein andFITC-CD28RE probe concentrations to be used in the FP assay. Therepresentative data for 10 nM and 0.33 nM are shown in FIG. 1A and FIG.1D, respectively. FIG. 1B shows results from cold competition withspecific and non-specific oligos. FIG. 1C shows the distribution of FPsignals in a representative 384-well plate.

For the 10 nM and 0.33 nM FP assays shown in FIG. 1A and FIG. 1D,respectively, the maximal Signal to Background (S/B) ratio was in therange of 8-11, indicating a robust assay. The background value for DNAprobe alone was ˜20 mP and the signal for c-Rel-CD28RE reaction was 200mP.

Electrophoretic Mobility Shift Assay (EMSA)

DNA binding reaction (20 μL) was carried out in 1×DNA Binding Buffer (10mM Tris, 40 mM NaCl, 1 mM EDTA, 4% glycerol) with 10 nM c-Rel proteinand 0.5 ng phosphor-labeled CD28RE oligonucleotide for 10 minutes atroom temperature in 96-well plates. Test compounds, in serial dilutions,were added into each well, and further incubated for 15 minutes beforeloading onto native 5% polyacrylamide gel. Electrophoresis proceeded for2.5 hours at 160V. Radioactive signals were quantified usingPhospho-Imager. IC₅₀ of compounds of example 1 to 75 were determined byquantifying the intensity of Rel/NF-κB inhibition in EMSA usingphospho-imager. An example of EMSA data of the present invention isshown for compound 6 in FIG. 2.

NF-κB GFP Assay

NFκB/Jurkat/GFP transcriptional reporter cell line was obtained from SBISystem Biosciences. NF-κB/Jurkat/GFP™ Reporter cells (5×10⁵ cells) wereplated at a concentration of 1 million cells/ml into each well of a24-well plate. TNF-α (5 ng/ml) was added. Compounds were added viaserial dilutions to corresponding wells. After 24 hours, 100 μl of thecells were transferred to a well of a Costar® UV plate (96 well, No lid,w/ UV Transparent Flat Bottom, Corning, N.Y., Cat#3635) and theintensity of GFP fluorescence was measured (Excitation 485+/−20,Emission 528+/−20) in a Synergy™ HT Multi-Detection Microplate Reader(BioTech, Winooski, Vt.). The intensities of GFP measured were plottedagainst the amount of TNF-α.

Pharmacokinetic Study of Compounds in CD-1 Mice

The study duration was two weeks, including acclimation and in-lifeportions. Three male mice were studied per compound. 5 mg/kg of thecompound of interest was administered intravenously via the tail vein ina single dose. Body weight was determined before the first dose. Thedosage vehicle was comprised of 100% PEG 400 and 2 meq NaOH. Bloodsamples were collected at 0.0833, 0.25, 0.5, 2, 4, and 8 hourspost-dose. Plasma was generated from blood with K₂EDTA as theanticoagulant.

Example 4 EMSA and NF-κB GFP Assay Results

EMSA NF-KB GFP Compound IC50 (μM) IC50 (μM) 1 <2 <30 2 <2 <30 3 <2 <30 4<2 <30 5 <2 <30 6 <1 <10 7 <2 <30 8 <0.5 <30 9 <5 <20 10 <1 <30 11 <1<20 12 <1 7 13 <1 9 14 <1 8.2 15 <0.5 <30 16 <0.5 <30 17 <0.5 <30 18<0.5 10 19 <1 <5 20 <1 <5 21 <1 <5 22 <1 <10 23 <1 <5 24 <1 <30 25 <1<30 26 <1 <30 27 <0.5 <5 28 <0.5 <5 29 <0.5 <5 30 <0.5 <10 31 <0.5 <5 32<0.5 <5 33 <0.5 <3 34 <0.5 10 35 <1 <10 36 <0.5 <10 37 <0.5 <20 38 <0.5<20 39 <0.5 <3 40 <0.5 5 41 <0.5 <10 42 <0.5 <10 43 <0.5 <3 44 <0.5 <345 <0.5 <5 46 <0.5 <5 47 <1 <5 48 <0.5 <5 49 <1 <5 50 <1 <5 51 <1 <5 52<0.5 <5 53 <5 <20 54 <5 <20 55 <5 <20 56 <5 <20 57 <5 <20 58 <5 <20 59<5 <20 60 <5 <20 61 <1 <5 62 <1 <5 63 <1 <5 64 <1 <5 65 <1 <5 66 <1 <567 <1 <5 68 <1 <5 69 <1 <5 70 <1 <5 71 <1 <5 72 <1 <5 73 <1 <5 74 <1 <575 <1 <5

Example 5 Pharmacokinetic Analysis

Pharmacokinetic parameters for compounds 13, 20, 26, 42, 44, and 46 areshown in FIGS. 3-8, respectively.

DOCUMENTS

-   1. Aly, H. M. and Kamal, M. M. (2012). Efficient one-pot preparation    of novel fused chromeno[2,3-d]pyrimidine and pryano[2,3-d]pyrimidine    derivatives. European Journal of Medicinal Chemistry 47: 18-23.-   2. Blythin, D. J., Domalski, M. S., Kim, Y. C., Kuo, J., and Liu,    J.-H. (1981). Simple synthetic route to “Oxa-Deaza-Flavins”    (2H-[1]-Benzopyrano [2,3-d] Pyrimidine-2,4 (3H)-Diones).    Heterocycles 16(2): 203-207.-   3. U.S. Pat. No. 4,272,535.-   4. International Application No. PCT/US2007/074233.

All documents cited in this application are hereby incorporated byreference as if recited in full herein.

Although illustrative embodiments of the present invention have beendescribed herein, it should be understood that the invention is notlimited to those described, and that various other changes ormodifications may be made by one skilled in the art without departingfrom the scope or spirit of the invention.

What is claimed is:
 1. A compound having the structure of formula (I):

wherein: A, B, C, and D are independently selected from the groupconsisting of carbon and nitrogen; X, Y, and Z are independentlyselected from the group consisting of oxygen, sulfur, and NR^(a); R₁ isselected from the group consisting of no atom, hydrogen, halogen, C₁₋₉alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH,—OR^(d), —OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a),—O(C═O)R^(d), —O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃,—CHF₂, —CH₂F, —CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a),—CONHCONR^(b)R^(a), —NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH,—CSR^(a), —CSOR^(a), —CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a),—S(C═O)R^(a), —S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R₂ is selected from thegroup consisting of no atom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH, —OR^(d),—OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d),—O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F,—CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a),—NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a),—CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a),—S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R₃ and R₄ are independently selectedfrom the group consisting of no atom, hydrogen, halogen, C₁₋₉ alkyl,C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH, —OR^(d),—OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d),—O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F,—CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a),—NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a),—CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a),—S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R₅ is selected from the groupconsisting of hydrogen, C₅₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl,C₁₋₉ heterocyclic, —R^(a)CO, —R^(a)NHCO, and R^(a)OCO; R₆ is selectedfrom the group consisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₂₋₉alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic; R^(a) is selectedfrom the group consisting of hydrogen, amine, C₁₋₉ alkyl, C₂₋₉ alkenyl,C₂₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic; R^(b) and R^(c) areindependently selected from the group consisting of oxygen, amine, C₁₋₉alkylene, C₂₋₉ alkenylene, C₂₋₉ alkynylene, arylene, and C₁₋₉heterocyclic; and R^(d) is selected from the group consisting ofhydrogen, amine, C₂₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉heterocyclic; wherein: if X is sulfur, R₂ is selected from the groupconsisting of no atom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉alkynyl, aryl, C₁₋₉ heterocyclic, —OR^(d), —OR^(b)OR^(a),—OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d), —O(C═O)OR^(a),—O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH,—COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a), —NR^(b)R^(a),—NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(b)R^(a),—CSNHCSNR^(b)R^(a), —SH, —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(b)R^(a); and, R^(d) is selected from the group consisting ofamine, C₂₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉heterocyclic; or a hydrate or pharmaceutically acceptable salt thereof.2. A compound according to claim 1, wherein: X, Y, and Z areindependently selected from the group consisting of oxygen and sulfur;R₁ is selected from the group consisting of —H, —F, —Cl, —Br, and —OEt;R₂ is selected from the group consisting of —H, —CH₃, —OH, —OEt, -Me,-Et, -nPr, —O-nPr, -OEtnPr, —OC₄H₉, —OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅,—O-isobutyl, —O-isopentyl, —OC_(n)H_(2n)OMe,—OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

R₃ is selected from the group consisting of —H, —Cl, —Br, and —F; R₄ is—H; R₅ is selected from the group consisting of —H and -Ph; and R₆ isselected from the group consisting of —H and —CH₃, m is 2, 3, 4 or 5; nis 2, 3, 4, or 5; wherein: if X is sulfur, R₂ is selected from the groupconsisting of —H, —CH₃, —OEt, -Me, -Et, -nPr, —O-nPr, —OEtnPr, —OC₄H₉,—OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅, —O-isobutyl, —O-isopentyl, —OC_(n)H_(2n)OMe,—OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

or a hydrate or pharmaceutically acceptable salt thereof.
 3. A compoundaccording to claim 1, wherein: X, Y, and Z are independently selectedfrom the group consisting of oxygen and sulfur; and R₆ is hydrogen,wherein: if X is sulfur, R₂ is selected from the group consisting of noatom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl,C₁₋₉ heterocyclic, —OR^(d), —OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a),—OR^(b)(C═O)R^(a), —O(C═O)R^(d), —O(C═O)OR^(a), —O(C═O)NR^(b)R^(a),cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH, —COR^(a), —COOR^(a),—CONR^(b)R^(a), —CONHCONR^(b)R^(a), —NR^(b)R^(a), —NHCOR^(a),—NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(b)R^(a),—CSNHCSNR^(b)R^(a), —SH, —S(C═O)R^(a), —S(C═O)OR^(a), S(C═O)NR^(b)R^(a);and, R^(d) is selected from the group consisting of amine, C₂₋₉ alkyl,C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic; or a hydrate orpharmaceutically acceptable salt thereof.
 4. A compound according toclaim 3, wherein: Z is oxygen; R₁ and R₃ are selected from the groupconsisting of hydrogen, halogen, —CN, and —CF₃; R₂ is selected from thegroup consisting of C₂₋₉ alkoxy, —OC_(n)H_(2n)OMe,—OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH₂,—O—C_(n)H_(2n)CONHMe, and —OH; m is 2, 3, 4 or 5; n is 2, 3, 4, or 5;and R₄ is selected from the group consisting of hydrogen, C₂₋₉ alkoxyand —OH, or a hydrate or pharmaceutically acceptable salt thereof.
 5. Acompound according to claim 4, wherein: X and Y are oxygen; and R₄ ishydrogen, or a hydrate or pharmaceutically acceptable salt thereof.
 6. Acompound according to claim 5, wherein the compound is selected from thegroup consisting of:

and hydrates or pharmaceutically acceptable salts thereof.
 7. A compoundselected from the group consisting of:

and hydrates or pharmaceutically acceptable salts thereof.
 8. A compoundselected from the group consisting of:

and hydrates or pharmaceutically acceptable salts thereof.
 9. A compoundcapable of inhibiting NF-κB, having the structure of formula (I):

wherein: A, B, C, and D are independently selected from the groupconsisting of carbon and nitrogen; X, Y, and Z are independentlyselected from the group consisting of oxygen, sulfur, and NR^(a); R₁ isselected from the group consisting of no atom, hydrogen, halogen, C₁₋₉alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH,—OR^(d), —OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a),—O(C═O)R^(d), —O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃,—CHF₂, —CH₂F, —CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a),—CONHCONR^(b)R^(a), —NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH,—CSR^(a), —CSOR^(a), —CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a),—S(C═O)R^(a), —S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R₂ is selected from thegroup consisting of no atom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH, —OR^(d),—OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d),—O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F,—CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a),—NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a),—CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a),—S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R₃ and R₄ are independently selectedfrom the group consisting of no atom, hydrogen, halogen, C₁₋₉ alkyl,C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, C₁₋₉ heterocyclic, —OH, —OR^(d),—OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d),—O(C═O)OR^(a), —O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F,—CHO, —COOH, —COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a),—NR^(b)R^(a), —NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a),—CSNR^(b)R^(a), —CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a),—S(C═O)OR^(a), —S(C═O)NR^(b)R^(a); R⁵ is selected from the groupconsisting of hydrogen, C₅₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl,C₁₋₉ heterocyclic, —R^(a)CO, —R^(a)NHCO, and —R^(a)OCO; R₆ is selectedfrom the group consisting of hydrogen, hydroxyl, amine, C₁₋₉ alkyl, C₂₋₉alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic; R^(a) is selectedfrom the group consisting of hydrogen, amine, C₁₋₉ alkyl, C₂₋₉ alkenyl,C₂₋₉ alkynyl, aryl, and C₁₋₉ heterocyclic; R^(b) and R^(c) areindependently selected from the group consisting of oxygen, amine, C₁₋₉alkylene, C₂₋₉ alkenylene, C₂₋₉ alkynylene, arylene, and C₁₋₉heterocyclic; and R^(d) is selected from the group consisting ofhydrogen, amine, C₂₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉heterocyclic; wherein: if X is sulfur, R₂ is selected from the groupconsisting of no atom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉alkynyl, aryl, C₁₋₉ heterocyclic, —OR^(d), —OR^(b)OR^(a),—OR^(c)OR^(b)OR^(a), —OR^(b)(C═O)R^(a), —O(C═O)R^(d), —O(C═O)OR^(a),—O(C═O)NR^(b)R^(a), cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH,—COR^(a), —COOR^(a), —CONR^(b)R^(a), —CONHCONR^(b)R^(a), —NR^(b)R^(a),—NHCOR^(a), —NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(b)R^(a),—CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(b)R^(a); and, R^(d) is selected from the group consisting ofamine, C₂₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉heterocyclic; or a hydrate or pharmaceutically acceptable salt thereof,wherein the compound inhibits NF-κB DNA-binding activity.
 10. A compoundaccording to claim 9, wherein: X, Y, and Z are independently selectedfrom the group consisting of oxygen and sulfur; R₁ is selected from thegroup consisting of —H, —F, —Cl, —Br, and —OEt; R₂ is selected from thegroup consisting of —H, —CH₃, —OH, —OEt, -Et, -nPr, —O-nPr, -OEtnPr,—OC₄H₉, —OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅, —O-isobutyl, —O-isopentyl,—OC_(n)H_(2n)OMe, —OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

R₃ is selected from the group consisting of —H, —Cl, —Br, and —F; R₄ is—H; R₅ is —H; and R₆ is selected from the group consisting of —H and—CH₃, m is 2, 3, 4 or 5; n is 2, 3, 4, or 5; wherein: if X is sulfur, R₂is selected from the group consisting of —H, —CH₃, —OEt, -Me, -Et, -nPr,—O-nPr, —OEtnPr, —OC₄H₉, —OC₅H₁₁, —OC₆H₁₃, —OC₇H₁₅, —O-isobutyl,—O-isopentyl, —OC_(n)H_(2n)OMe, —OC_(n)H_(2n)OC_(m)H_(2m)OMe,—OC_(n)H_(2n)OH, —OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, —O—C_(n)H_(2n)COOH, —O—C_(n)H_(2n)CONH2,—O—C_(n)H_(2n)CONHMe,

or a hydrate or pharmaceutically acceptable salt thereof.
 11. A compoundaccording to claim 9, wherein: X, Y, and Z are independently selectedfrom the group consisting of oxygen and sulfur; and R₆ is hydrogen,wherein: if X is sulfur, R₂ is selected from the group consisting of noatom, hydrogen, halogen, C₁₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl,C₁₋₉ heterocyclic, —OR^(d), —OR^(b)OR^(a), —OR^(c)OR^(b)OR^(a),—OR^(b)(C═O)R^(a), —O(C═O)R^(d), —O(C═O)OR^(a), —O(C═O)NR^(b)R^(a),cyano, nitro, —CF₃, —CHF₂, —CH₂F, —CHO, —COOH, —COR^(a), —COOR^(a),—CONR^(b)R^(a), —CONHCONR^(b)R^(a), —NR^(b)R^(a), —NHCOR^(a),—NR^(b)COR^(a), —CSOH, —CSR^(a), —CSOR^(a), —CSNR^(b)R^(a),—CSNHCSNR^(b)R^(a), —SH, —SR^(a), —S(C═O)R^(a), —S(C═O)OR^(a),—S(C═O)NR^(b)R^(a); and, R^(d) is selected from the group consisting ofamine, C₂₋₉ alkyl, C₂₋₉ alkenyl, C₂₋₉ alkynyl, aryl, and C₁₋₉heterocyclic; or a hydrate or pharmaceutically acceptable salt thereof.12. A compound according to claim 11, wherein: Z is oxygen; R₁ and R₃are selected from the group consisting of hydrogen, halogen, —CN, and—CF₃; R₂ is selected from the group consisting of C₂₋₉ alkoxy,—OC_(n)H_(2n)OMe, —OC_(n)H_(2n)OC_(m)H_(2m)OMe, —OC_(n)H_(2n)OH,—OC_(n)H_(2n)OC_(m)H_(2m)OH, —OC_(n)H_(2n)OEt,—OC_(n)H_(2n)OC_(m)H_(2m)OEt, and —OH; m is 2, 3, 4 or 5; n is 2, 3, 4,or 5; R₄ is selected from the group consisting of hydrogen, C₂₋₉ alkoxyand —OH, or a hydrate or pharmaceutically acceptable salt thereof.
 13. Acompound according to claim 12, wherein: X and Y are oxygen; and R₄ ishydrogen or a hydrate or pharmaceutically acceptable salt thereof.
 14. Acompound according to claim 13, wherein the compound is selected fromthe group consisting of:

and hydrates or pharmaceutically acceptable salts thereof.
 15. Acompound capable of inhibiting NF-κB, selected from the group consistingof:

and hydrates or pharmaceutically acceptable salts thereof, wherein thecompound inhibits NF-κB DNA-binding activity.
 16. A compound capable ofinhibiting NF-κB, selected from the group consisting of:

and hydrates or pharmaceutically acceptable salts thereof, wherein thecompound inhibits NF-κB DNA-binding activity.
 17. A pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound according to claim
 1. 18. The pharmaceutical composition ofclaim 17, wherein the compound is selected from the group consisting of:

and hydrates or pharmaceutically acceptable salts thereof.
 19. A methodof inhibiting NF-κB in a cell, comprising: contacting the cell with acompound according to claim
 1. 20. The method of claim 19, wherein thecompound is selected from the group consisting of:

and hydrates or pharmaceutically acceptable salts thereof.